Benzodiazepine derivative

ABSTRACT

A benzodiazepine derivative of the formula (I): ##STR1## wherein R 1  is a bond, --CH 2  --, --CH 2  O--, --SCH 2  -- or a group of the formula: ##STR2## R 2  is a lower alkyl, --COOR 5 , --CONH(CH 2 ) n  COOR 5 , --CONHSO 2  R 5 , --SO 2  NHCOR 5 , or an optionally substituted heterocyclic group (R 5  is a hydrogen atom, lower alkyl or benzyl and n is an integer of 1 to 5); R 3  is a bond, --CO-- or --CONH--; and R 4  is an optionally substituted heterocyclic group, optionally substituted lower alkyl, optionally substituted lower cycloalkyl, optionally substituted aryl, or lower alkoxycarbonyl group, or a pharmaceutically acceptable salt thereof, which has a high affinity for gastrin receptors and/or CCK-B receptors but not for CCK-A receptors, and is useful for treating diseases associated with gastrin receptors and/or CCK-B receptors without inducing the side effects associated with CCK-A receptors.

FIELD OF THE INVENTION

The present invention relates to novel benzodiazepine derivativescapable of competing with gastrin and/or CCK-B and binding to theirreceptors, and pharmaceutical compositions which contain the same andare useful in the treatment of various diseases associated with gastrinand/or CCK-B receptors.

BACKGROUND OF THE INVENTION

Gastrin and cholecystokinin (CCK) are physiologically active substancesbelonging to what is called a gastrin sub-family of the gastrointestinalpeptide hormone family. Although gastrin receptors are commonly found invarious tissues including the whole superior digestive tract, pancreas,liver, biliary duct and the like, they mainly exist on parietal cells offundic glands and participate in the mediation of gastric acidsecretion. As for CCK receptors, it is known that there are two types ofreceptors, i.e., CCK-A receptor found in peripheral tissues such asdigestive gut and CCK-B receptor found in brain. The former participatesin the control of gut motility and pancreas secretion whereas the latterin the control of central nervous action, appestat and the like.Accordingly, it has been expected that compounds capable of competingwith gastrin and/or CCK-B and binding to their receptors are useful inthe treatment of animals including human suffering from gastrointestinaland central nervous diseases associated with receptors for these peptidehormones. For example, such compounds are thought to be useful as ananti-tumor agent; a drug for treating pancreatitis, gallbladder disorderor irritable bowel syndrome, for relieving biliary colic, and forimproving appetite. Further, investigations into receptors in bothgastrointestinal and central nervous system revealed that thesegastrointestinal peptide hormones are also important as biologicallyactive substances "Brain and Peptides" Taisha, vol. 18, No. 10, 33-44(1981); J. Hughus, C. Woodruff, D. Horwell, A. McKnight & D. Hill,"Gastrin", J. H. Walsh ed., Rovan Press, Ltd., New York, 1993, p.169-186; F. Makovec, Drugs of the Future, 18, 919 (1993); JapanesePatent Publication (KOKAI) 63-238069, EP 167,919; U.S. Pat. No.4,820,834; EP 284,256; U.S. Pat. No. 5,004,741!.

For instance, gastrin antagonists specific to gastrin receptors arethought to be effective on gastrin-associated disorders such as pepticulcers in gaster and duodenum, Zollinger-Ellison syndrome, hyperplasiaof sinus C cells, and decrease in gastrin activity. The usefulness ofantagonists specific to gastrin-receptor in the treatment of gastric andduodenal ulcers has been reported (Taisha, 29/7, 1992, R. Eissele, H.Patberg, H. Koop, W.

Krack, W. Lorenz, A. T. McKnight & R. Arnold, Gastroenterology, 103,1596 (1992), etc.)

There have been reported that antagonists against CCK-B receptor areuseful in the reinforcement and elongation of the analgetic effect ofopioid-type compounds (e.g., morphine derivatives such as morphinesulfate or hydrochloride) which. are antagonistic against opioidreceptors Drugs of the future 18, 919 (1993); Proc. Natl. Acad. Sci.USA, Vol. 87, p. 71, 05 September 1990, Neurobiology!.

It is necessary to use a compound capable of binding to an intendedpeptide hormone receptor in preference discriminating it from that forpeptide hormones of different sub-types in order to conduct treatmentmore efficiently.

A series of benzodiazepine derivatives which are antagonistic againstgastrin or CCK-B receptor have been disclosed (WO 93/14074 and WO93/14075). However, they failed to disclose any specific pharmacologicaldata regarding antagonistic activity against gastrin receptors, orantagonists useful as a medicine. Further, all the compounds disclosedin these publications are racemates with an asymmetric carbon atom atthe 3-position, which makes the preparation thereof difficult andrequires optical resolution to obtain a single compound.

Accordingly, it has been strongly demanded to develop a compound whichcan bind to an intended receptor discriminating it from other peptidehormone receptor, and is useful as a drug and producible in ease.

DISCLOSURE OF THE INVENTION

In the situations above, the present inventors have studied intensivelyto develop compounds which have high affinity for gastrin receptorsand/or CCK-B receptors with high selectivity but low or no affinity forCCK-A receptors, and found that certain benzodiazepine derivatives areuseful for the purposes above and established the present invention.

Thus, the present invention provides a compound of the formula (I):##STR3## wherein R₁ is a bond, --CH₂ --, --CH₂ O--, --OCH₂ --, --SCH₂ --or a group of the formula: ##STR4## R₂ is a lower alkyl, --COOR₅,--CONH(CH₂)_(n) COOR₅, --CONHSO₂ R₅, --SO₂ NHCOR₅, or an optionallysubstituted heterocyclic group (R₅ is a hydrogen atom, lower alkyl orbenzyl and n is an integer of 1 to 5); R₃ is a bond, --CO-- or --CONH--;and R4 is an optionally substituted heterocyclic group, optionallysubstituted lower alkyl, optionally substituted lower cycloalkyl,optionally substituted aryl, or lower alkoxycarbonyl group, or apharmaceutically acceptable salt thereof.

Although all the compounds (I) as defined above are useful to achievethe purposes of the present invention, those of the formula (I) whereinR₃ is --CO-- and R₄ is a lower cycloalkyl group and/or R₁ --R₂ is--COOR₅, --CONHSO₂ R₅, --SO₂ NHCOR₅, --CH₂ COOR₅, --OCH₂ COOR₅, --SCH₂COOR₅, tetrazolylmethyloxy or a 5-membered heterocyclic group containingan N atom are preferable.

THE BEST EMBODIMENT FOR PRACTICING THE INVENTION

Throughout the present specification, the terms "gastrin receptorantagonist" or "CCK-B receptor antagonist" is referred to a compoundcapable of competitively inhibiting the binding of gastrin receptor orCCK-B receptor with respective natural ligand, and is usedinterchangeably with the term "gastrin antagonist" or "CCK-Bantagonist", respectively. Since the compound (I) of the presentinvention has a strong affinity for gastrin receptors and/or CCK-Breceptors and can bind to them specifically competing with their naturalligands, it may be referred to as "gastrin receptor antagonist" or"CCK-B receptor antagonist".

Because of the same reason above, the terms "gastrin receptorantagonism" and "gastrin antagonism", and the terms "CCK-B receptorantagonism" and "CCK-B antagonism" are used exchangeably.

The following terms used in the definition of compound (I) are definedbelow.

The term "lower alkyl" means straight or branched chain C_(1-C) ₈hydrocarbon group including methyl, ethyl, n-propyl, i-propyl, n-butyl,s-butyl, t-butyl, n-pentyl, i-pentyl, neopentyl, s-pentyl, t-pentyl,n-hexyl, neohexyl, i-hexyl, s-hexyl, t-hexyl, heptyl and octyl, and C₁-C₃ hydrocarbon group is preferred.

The term "lower cycloalkyl" means C₃ -C₇ cycloalkyl group includingcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, and C₃-C5 cycloalkyl is preferred.

The term "heterocyclic group" means 5- to 7-membered both aromatic- andnon-aromatic heterocyclic groups containing one or more hetero atomsselected independently from the group consisting of O, S and N. Examplesof aromatic heterocyclic group include furyl, thienyl, tetrazolyl,pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, pyridinyl,oxadinyl and triazinyl. Examples of non-aromatic heterocyclic groupinclude pyrrolidinyl, thiazolidinyl, oxazolidinyl, imidazolidinyl,thiazolinyl, oxazolinyl, imidazolinyl, piperidinyl, piperadinyl,morpholinyl, thiomorpholinyl, oxadiazolyl and dioxanyl.

Preferred heterocyclic groups are pyrrolidinyl, thiazolidinyl andthienyl for R₄, and tetrazolyl, 5-keto-1,2,4-oxadiazolyl and the likefor R₂.

The term "aryl" means phenyl, naphthyl and the like.

Examples of substituents in R₂ or R₄ are hydroxy, carbonyl, aminooptionally protected with an amino-protecting group, halogen (F, Cl, Br,etc.), lower alkyl and lower alkoxy.

The term "lower alkoxy" means straight or branched chain C₁ -C₆ alkoxygroup including methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy,s-butoxy, t-butoxy, n-pentyloxy, i-pentyloxy, neopentyloxy, s-pentyloxy,t-pentyloxy, n-hexyloxy, neohexyloxy, i-hexyloxy, s-hexyloxy andt-hexyloxy, and C₁ -C₃ alkoxy is preferred.

The benzodiazepine derivatives (I) of the present invention are noveland can be prepared, for example, by alkylating a compound (IV)(3-amino-1H-1,5-benzodiazepine-2,4-(3H,5H)-dione), at the 1- and5-positions and 3-amino group as described below.

As can be seen from the formula (I), the benzodiazepine derivatives ofthe present invention have a plane-symmetric structure and are notracemates. This advantageous feature facilitates the preparation of anintended single compound without optical resolution.

The compounds (I) of the present invention can be prepared using anymethods known in the art as exemplified below. The following processesare, however, provided for the illustrative purpose only and the scopeof the invention should not be limited to compounds (I) preparedaccording to these processes.

Method 1

A benzodiazepine compound of the formula (IV): ##STR5## is reacted witha compound of the formula (II): ##STR6## wherein R₁ and R₂ are asdefined above to form a compound of the formula (III) having an ureabond at the 3-position: ##STR7## wherein R₁ and R₂ are as defined above.The compound (III) is then reacted with a compound of the formula (V):

    R.sub.4 -R.sub.3 --CH.sub.2 X                              (V)

wherein X is a halogen, and R₃ and R₄ are as defined above forN-alkylation at 1- and 5-positions to give the objective compound (I).

Method 2

An benzodiazepine (IV) is protected at the 3-amino group and N-alkylatedby reacting with a compound (V) in a manner similar to that described inMethod 1 above, which is followed by deprotection, reaction with acompound (II), and alkylation at the 3-position to give the objectivecompound (I).

The preparation of compounds (I) of the present invention will beexplained in more detail below referring to the method 1. The method 2can be effected in substantially the same manner except for additionalamino-protection and deprotection procedures.

The starting material, i.e.,3-amino-1H-1,5-benzodiazepine-2,4-(3H,5H)-dione (IV), is a knowncompound and is prepared by any methods described in literatures orthose known in the art.

The said compound (IV) is reacted with a compound (II) under theconditions for alkylation, generally in a solvent such asdimethylformamide, methylene chloride or the like at room temperaturefor about 0.5 to 2 hr.

The N-alkylation of a compound (III) is carried out in general byreacting a compound (III) with a halide of the formula R₄ -R₃ --CH₂ X(V) in the presence of a base such as potassium carbonate or the likeand a salt such as potassium iodide or the like in a solvent such asdimethylformamide or the like at room temperature for about 10 to 20 hr.Conventional reagents for N-alkylation such as KOH and (n-Bu)₄ N₊ Br⁻,NaH, t-BuOK, NaNH₂ and the like are also usable.

A compound of the formula (I) produced by the processes above can befurther converted into compounds which are also shown by the formula (I)through the hydrolysis with lithium hydroxide or the like in analcoholic solvent or the like. The purification of a final product canbe carried out in a conventional manner, for example, extracting with anorganic solvent such as ethyl acetate, drying, concentrating and/orchromatographing.

A compound of the formula (I) can be optionally converted into apharmaceutically acceptable salt using an appropriate method known inthe art.

The compound (I) of the present invention forms a salt with conventionalinorganic or organic acids, or inorganic or organic bases. Examples ofsalts of the compound (I) include those formed with alkali metals suchas sodium, potassium and the like, alkali earth metals such as calcium,magnesium and the like; organic bases such as ammonium, trimethylamine,triethylamine, pyridine, picoline, dicyclohexylamine,N,N'-dibenzylethylenediamine and the like; organic acids such as aceticacid, maleic acid, tartaric acid, methanesulfonic acid, benzenesulfonicacid, formic acid, toluenesulfonic acid, trifluoroacetic acid and thelike; inorganic acids such as hydrochloric acid, hydrobromic acid,sulfuric acid, phosphoric acid and the like; and amino acids such asarginine, aspartic acid, glutamic acid and the like.

An intermediate of the formula (III) described above is a novel compoundwhich is useful in the synthesis of not only the compound (I) of thepresent invention but also various other compounds. The compound (III)can be prepared by any methods known in the art in addition to thosedescribed above. For instance, it can be prepared according to thereaction schemes below. All the compounds of the formula (III) areuseful as intermediates for the preparation of the compound (I) of thepresent invention irrespective of the process of preparation. ##STR8##

An in vivo experiment (Schild method, Experiment 1 below) revealed thatthe compound (I) of the present invention can inhibit the gastric acidsecretion. As is shown in Experiment 2 below, in vitro experiments havebeen conducted to examine gastrin- or CCK-B-receptor antagonism ofvarious compounds (I), which revealed that the benzodiazepinederivatives of the present invention have the activities of both types.The experimental results obviously indicate that the benzodiazepinederivatives of the present invention inhibit the gastric acid secretionand are antagonistic against gastrin receptors and/or CCK-B receptorswhile well discriminating them from CCK-A receptors.

Accordingly, the present invention provides a pharmaceutical compositioncomprising therapeutically effective amount of a compound (I) inassociation with pharmaceutically acceptable carriers therefor, which isuseful in the treatment of diseases caused by physiological disordersnormally controlled through gastrin receptors, for example, gastriculcer, duodenal ulcer, gastritis, reflux esophagitis andZollinger-Ellison syndrome, without inducing any side effects associatedwith CCK-A receptors.

The present invention also provides a pharmaceutical compositioncomprising therapeutically effective amount of a compound (I) inassociation with pharmaceutically acceptable carriers therefor, which isuseful as an antianxiety drug, or in the treatment of central nervousdisorders induced by physiological disorders normally controlled throughCCK-B receptors, for example, diseases caused by disorder ofappestat-control-system without side effects associated with CCK-Areceptors. Further, because the compound (I) seems to be able to enforceor lengthen the analgetic effect induces by opioid-type drugs, it isusable in combination with such analgesics.

The compound (I) of the present invention may be used alone or incombination with one or more other drugs. The combination treatment canbe carried out in a manner known in the art by administering a compound(I) and one or more pharmaceutically active ingredients as a singlecomposition, or successively .

The compound (I) of the present invention, when used in combination withan existing drug commonly used as an anti-peptic ulcer agent such ashistamine H₂ blocker (H₂ B) including cimetidine (Smithkline Beecham),ranitidine (GLAXO), roxatidine (Teikokuzouki), famotidine (Yamanouchi)and the like, or proton-pump inhibitors including omeprazole(Yamanouchi), advantageously exerts the anti-ulcer activity whilesuppressing the side effects of the co-existing drug through theinherent gastrin inhibitory effect. Thus, one of remarkable drawbacks ofH₂ B and proton-pump inhibitors is the high incidence of post-treatmentrelapse following a chronic administration. There are two factors knownto be involved in the relapse of ulcer following the H₂ B treatment,i.e., (1) rebound phenomenon of acid secretion; and (2) the decrease inthe protective function of gastric mucosa. The hypergastrinemia due tochronic administration of a proton-pump inhibitor is also related to theulcer relapse. The present inventors demonstrated that such reverseeffect of H₂ B or proton-pump inhibitor could be prevented byadministering it in combination with a gastrin receptor antagonist (see,Experiment 4 below). In the Experiment 4, a combined formulation (drug)of a typical H₂ B (famotidine), and a known gastrin receptor antagonist(L-365,260 described in Example 281 of Japanese Patent Publication(KOKAI) 63-238069 (EP 167,919; EP 284,256; U.S. Pat. No. 4,820,834; U.S.Pat. No. 5,004,741)) was used to evaluate the inhibitory effect ofL-365,260 on the relapse of ulcer following a continuous administrationof H₂ B on the basis of the appearance of phenomena (1) and (2) above.The result showed that L-365,260 suppressed the appearance of thephenomena (1) and (2) which generally accompanies to the famotidineadministration, demonstrating that it is possible to prevent the relapseof ulcer following the anti-ulcer treatment with H₂ B. This resultindicates that the compound (I) of the present invention having anactivity to antagonizing against gastrin receptor should be useful inthe prevention of relapse of peptic ulcer after continuousadministration of H₂ B such as famotidine.

The result obtained in Experiment 4 below also indicates that thecompound (I) of the present invention, owing to its activity, inhibitsthe hypergastrinemia following the continuous administration ofproton-pump inhibitors such as omeprazole and the like and is useful inthe prevention of relapse of ulcer following the treatment withproton-pump inhibitors. These results indicate that the compound of thepresent invention is useful in the treatment of refractory ulcers andcontribute to solve the problems associated with conventional anti-ulceragents.

Accordingly, the present invention also provides a pharmaceuticalcomposition for treating ulcers, which comprises a compound (I) of thepresent invention and an H₂ B or a proton-pump inhibitor in associationwith pharmaceutically acceptable carriers therefor.

Such a composition may contain a compound (I) and an H₂ B or proton-pumpinhibitor(s) in the ratio of 1-3:3-1, preferably 1:1.

A hybrid-type compound can be prepared by coupling an appropriate H₂ Binhibitor to a compound (I) at its R₁ -R₂ or R₃ -R₄ moiety as shown inReference Example below.

When using a compound (I) of the present invention in treatment, it canbe administered orally or parenterally. In the case of oraladministration, a compound of the present invention may be formulatedinto ordinary formulations in the form of solid such as tablets,powders, granules, capsules and the like; solutions; oily suspensions;liquid formulations such as syrups, elixirs and the like. In the case ofparenteral administration, a compound of the present invention may beformulated into an aqueous or oily suspension for injection. Inpreparing the formulations, conventional excipients, binders,lubricants, aqueous solvents, oily solvents, emulsifiers, suspendingagents or the like may be used. The formulations may contain otheradditives, such as preservatives, stabilizers or the like.

Although appropriate daily dosage of the compound of the presentinvention varies depending on the administration route, age, bodyweight, conditions of the patient, and the kind of disease to betreated, in the case of adult patients, it can generally be betweenabout 10-200 mg, preferably about 20-100 mg on oral administration, andabout 1-20 mg, preferably about 2-10 mg on parenteral administration, in1-2 divisions.

When administering the compound of the present invention as a combinedformulation, the dose will be determined on the basis of the doseindicated above.

The following Examples are provided to further illustrate the presentinvention and are not to be construed as limiting the scope thereof.##STR9##

Preparation 1 Diethyl methoxyiminomalonate 2

A solution of diethyl ketomalonate 1 (51.0 g, 0.293 mmol),o-methylhydroxylamine hydrochloride (24.46 g, 0.293 mmol) and pyridine(23.2 g, 0.293 mmol) in ethanol (250 ml) is heated to reflux for 3 hr.The solvent is removed under reduced pressure. The residue is dissolvedin ethyl acetate and washed with water, dilute hydrogen chloride,aqueous sodium hydrogencarbonate solution, and water successively. Afterdrying over sodium sulfate, the solvent is removed under reducedpressure. The resultant residue is distilled under reduced pressure at80°-85° C./0.5 mmHg to obtain Compound 2 (55.5 g, 93.2%). NNMR (CDCl₃)δ:1.35 (3H, t, J=7.1 Hz), 1.35 (3H, t, J=7.1 Hz), 4.11 (3H, s), 4.36 (2H,q, J=7.1 Hz), 4.37 (2H, q, J=7.1 Hz).

Preparation 2 3-Methoxyimino-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 3

A mixture of iN sodium methylate (162 ml), o-phenylenediamine (17.5 g,162 mmol) and diethyl 2-(methoxyimino)malonate (32.91 g, 162 mmol) isheated to reflux for 5 hr. After cooling, the mixture is acidified with2N HCl (162 ml) and pale yellowish crystals (14.3 g, 41%) are filteredoff.

IR ν_(max) (nujol): 1699, 1655, 1460, 1375 cm¹. NMR (CDCl₃ +CD₃ OD)8:4.03 (3H, s), 7.10-7.28 (4H, m).

Preparation 3 3-Amino-1H-1,5-benzodiazepine-2,4(3H,5H )-dione 4

A solution of 3-methoxyimino-1H-1,5-benzodiazepine-2,4(3H,5H)-dione(3.78 g, 17.4 mmol) and 10% Pd/C (1.8 g) in methanol (300 ml) is stirredfor 15 hr under hydrogen gas. After removing the catalyst by filtration,the filtrate is concentrated under reduced pressure. The resultantresidue is crystallized from methanol to obtain Compound 4 (2.062 g,62%). M.p. =290°-291° C.

IR ν_(max) (nujol): 3376, 3287, 1704, 1673, 1563 cm⁻¹.

NMR (DMSO-d₆)6: 3.75 (1H, s), 7.09-7.25 (4H, m).

Elemental Analysis (for C₉ H₉ N₃ O₂ ·0.1H₂ O)

Found: C, 56.16;H, 4.88; N, 21.64

Calcd.: C, 56.01;H, 4.80; N, 21.77.

Preparation 43-(N'-(m-tolyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 5

A mixture of 3-amino-1H-1,5-benzodiazepine-2,4(3H,5H)-dione (1.761 g,9.21 mmol) and m-tolylisocyanate (1.31 g, 10.13 mmol) indimethylformamide (17 ml) is stirred for 1 hr under ice-cooling. To thereaction mixture is added diisopropyl ether (50 ml) and the resultantcrystalline precipitates are filtered off to obtain Compound 5 (2.98 g;yield, 99%). M.p. >300° C.

IR v max (nujol): 3350, 3301, 3215, 3072, 1714, 1656, 1600, 1562 cm⁻¹.

NMR (DMSO-d₆)δ: 2.23 (3H, s), 4.63 (1H, d, J=7.4 Hz), 6.73 (1H, d,J=7.0Hz), 6.82 (1H, d, J=7.4 Hz), 7.31-7.00 (8H, m), 10.77 (2H, s).

Elemental Analysis (for C₁₇ H₁₆ N₄ O₃ ·0.1H₂ O)

Found: C, 62.45; H, 5.03; N, 17.13

Calcd.: C, 62.61; H, 5.01; N, 17.18.

Preparation 53-(N'-(3-(3-(carbomethoxy)propyl-carbamoyloxymethyl)phenyl)ureido)--1H-1,5-benzodiazepine-2,4(3H,5H)-dione6

A mixture of 3-amino-1H-1,5-benzodiazepine-2,4(3H,5H)-dione (1.91 g,10.0 mmol) and3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenylisocyanate (3.507 g,12.0 mmol) in dimethylformamide (19 ml) is stirred for 1 hr at roomtemperature. To the residue obtained by concentrating the reactionmixture under reduced pressure is added methylene chloride (300 ml) andmethanol (100 ml), and the mixture is stirred for 30 min at roomtemperature. The reaction mixture is filtered through a silica gel (50g) layer to obtain Compound 6 (4.822 g, 100%) as colorless crudecrystals. M.p. =151°-155° C.

IR ν_(max) (nujol): 3273, 1722, 1698, 1636, 1599, 1567, 1529, 1502cm⁻¹.

NMR (DMSO-d₆)δ: 1.64 (2H, qui, J=7.0 Hz), 2.30 (2H, t, J=7.4 Hz), 3.00(2H, q, J=5.8 Hz), 3.57 (3H, s), 4.64 (1H, d, J=7.4 Hz), 4.93 (2H,s),6.79-6.92 (2H, m), 7.12-7.40 (8H, m), 9.18 (1H, s), 10.78 (2H,s).

Preparation 63-(N'-(3-(3-(benzyloxycarbonyl)propyl-carbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione7

Compound 7 is prepared in a manner similar to that used for preparationof Compound 6 above. M.p. =233°-235° C.

IR ν_(max) (KBr): 3386, 3287, 1716, 1697, 1636, 1599, 1566 cm⁻¹.

NMR (DMSO-d₆ 1)δ: 1.67 (2H, qui, J=7.2 Hz), 2.37 (2H, t, J=7.4 Hz), 3.01(2H, q, J=6.2 Hz), 4.64 (1H, d, J=7.6 Hz), 4.93 (2H, s), 5.07 (2H, s),6.86 (2H, dd, J=5.7, 7.1 Hz), 7.14-7.40 (8H, m), 7.36 (5H, s), 9.18 (1H,s), 10.77 (2H, S).

Elemental Analysis (for C₂₉ H₂₉ N₅ O₇ ·0.5H₂ O)

Found: C, 61.32; H, 5.37; N, 12.55

Calcd.: C, 61.26; H, 5.32; N, 12.32.

Preparation 7 Chloromethyl cyclopentyl ketone

To cyclopentanecarboxylic acid (5.71 g, 5 mmol) is added thionylchloride (11.9 g, 10 mmol) and the mixture is stirred for 2 hr at roomtemperature. After removing excessive thionyl chloride under reducedpressure, the mixture is distilled at 48-52/18 mmHg to obtain acidchloride (5.54 g, 83.5%). A solution of the acid chloride (5.54 g) inether (5 ml) is added dropwise to a solution of excessive diazomethanein ether under ice-cooling. The mixture is stirred for 30 min andconcentrated under reduced pressure to about half of its originalvolume. The solution is added dropwise to conc. HCl at -20° C. and themixture is stirred for 3 hr. After adding ice-cold water, the organiclayer is separated, washed with water and dried over sodium sulfate. Thesolvent is removed under reduced pressure and the residue is distilledat 88-92/18 mmHg to obtain the objective compound (3.53 g, 58.4%).

NMR (CDCl₃)δ: 1.5-2.0 (8H, m), 3.12 (1H, m), 4.17 (2H, s).

In a manner similar to that described above, 2-(chloroacethyl)furan,2-(chloroacethyl)thiophene, 4-(chloroacetyl)-1,2-dimethoxybenzene,chloroacetyl-cyclopropane, and o-methylphenacyl chloride were prepared.

Preparation 8 Bromoacetylpyrrolidine

A solution of pyrrolidine (3.97 g, 55 mmol) and triethylamine (5.84 g,57.7 mmol) in methylene chloride (25 ml) is added dropwise to a solutionof bromoacetyl bromide (11.28 g, 91.7 mmol) in methylene chloride (25ml) under ice-cooling. The mixture is stirred at 0° C. for 30 min thenat room temperature for 30 min, and poured into ice-cold water. Theorganic layer is separated, washed with water, dried over sodium sulfateand concentrated under reduced pressure.

In a manner similar to that described above, bromoacetylthiazolidine,cyclopropylchloroacetamide were prepared. ##STR10##

Preparation 93-(t-Butoxycarbonylamino)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 13

To a suspension of 3-amino-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 4(3.27 g, 17.1 mmol) in tetrahydrofuran (300 ml) is added di-tert-butyldicarbonate (5.62 g, 25.8 mmol). The mixture is stirred for 24 hr atroom temperature and concentrated. To the residue is added a mixture(200 ml) of methylene chloride/methanol (9:1) and water (50 ml) and themixture is stirred for 10 min. The organic layer is dried over magnesiumsulfate and concentrated under reduced pressure. Crystallization from amixture of methylene chloride/methanol and diisopropyl ether yields thetitled Compound 13 (4.5 g, Yield 90%) as white crystals. M.p. =243°-244°C.

NMR (DMSO-d₆)δ: 1.38 (9H, m), 4.51 (1H, d. J=8.2 Hz), 6.48 (1H, d. J=8.2Hz), 7.14-7.31 (4H, m), 10.54-10.93 (2H, broad).

Elemental Analysis (for C₁₄ H₁₇ N₃ O₄)

Calcd.: C, 57.72; H, 5.88; N, 14.43

Found: C, 57.45; H, 5.88; N, 14.36.

Preparation 101,5-Bis-(cyclopropylcarbonylmethyl)-3-(t-butoxycarbonylamino)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione14 h

A suspension of3-(t-butoxycarbonylamino)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 13(2.039 g, 7 mmol), cyclopropylcarbonylmethyl chloride (2.488 g, 21mmol), potassium carbonate (2.902 g, 21 mmol) and potassium iodide (174mg, 1.05 mmol) in dimethylformamide (20 ml) is stirred for 15 hr at roomtemperature. The reaction mixture is concentrated under reducedpressure. Purification of the residue by column chromatography on silicagel (toluene:ethyl acetate, 2:1) gives the titled Compound 14 h (3.188g; yield, 100%) as a foam.

IR ν_(max) (KBr): 3445, 1700, 1658, 1503, 1450, 1320 cm⁻¹.

NMR (DMSO-d₆)δ: 0.81-1.06 (8H, m) 1.35 (9H, s), 2.05-2.22 (2H, m), 4.82(2H, d, J=18.4 Hz), 4.85 (1H, d, J=8.2 Hz), 4.96 (2H, d, J=18.4 Hz),6.73 (1H, d, J=8.2 Hz), 7.27-7.45 (4H, m).

Elemental Analysis (for C₂₄ H₂₉ N₃ O₆ ·0.2H₂ O)

Calcd.: C, 62.79; H, 6.45; N, 9.15

Found: C, 62.92; H, 6.44; N, 8.94.

Preparation 111,5-Bis-(pyrrolidinocarbonylmethyl)-3-(t-butoxycarbonylamino)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 14a

Compound 14a is prepared in a manner similar to that used forpreparation of Compound 14 h using previously prepared Compound 13 andpyrrolidinocarbonylmethyl bromide.

M.p. =137°-139° C.

IR ν_(max) (KBr): 3440, 1700, 1503, 1420 cm⁻¹.

NMR (DMSO-d₆)δ: 1.36 (9H, s), 1.65-1.96 (8H, m), 3.23-3.38 (4H, m),3.39-3.52 (4H, m), 4.47 (2H, d, J=16.6 Hz), 4.68 (2H, d, J=16.6 Hz),4.83 (1H, d, J=8.2 Hz), 6.57 (1H, d, J=8.2 Hz), 7.32-7.46 (2H, m),7.47-7.57 (2H, m).

Elemental Analysis (for C₂₆ H₃₅ N₅ O₆ ·0.7H₂ O)

Calcd.: C, 59.35; H, 6.97; N, 13.31

Found: C, 59.35; H, 6.84; N, 13.14.

Preparation 121,5-Bis-(thienylcarbonylmethyl)-3-(t-butoxycarbonylamino)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione14g

Compound 14g is prepared in a manner similar to that used forpreparation of Compound 14 h using previously prepared Compound 13 and2-thienylcarbonylmethyl chloride.

M.p. =132°-135° C.

IR ν_(max) (KBr): 3435, 1703, 1672, 1503, 1419 cm⁻¹.

NMR (DMSO-d₆)δ: 1.36 (9H, s), 4.96 (1H, d, J=8.4 Hz), 5.27 (2H, d,J=18.0Hz), 5.52 (2H, d, J=18.0 Hz), 6.82 (1H, d, J=8.4 Hz), 7.28-7,36(2H, m), 7.45 (4H, d, J=2.0Hz), 8.10-8.18 (4H, m).

Elemental Analysis (for C₂₆ H₂₅ N₃ O₆ S₂ ·0.2H₂ O)

Calcd.: C, 57.49; H, 4.71; N, 7.74; S, 11.80

Found: C, 57.54; H, 4.81; N, 7.71; S, 11.69.

Preparation 13 1,5-Bis-(cyclopropylmethyl)-3-(t-butoxy-carbonylamino)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 14o

Compound 14o is prepared in a manner similar to that used forpreparation of Compound 14 h using previously prepared Compound 13 andcyclopropylmethyl chloride.

M.p. =156°-157° C.

IR ν_(max) (KBr): 3430, 3370, 1695, 1500, 1419 cm⁻¹.

NMR (DMSO-d₆) δ: 0.04-0.17 (4H, m), 0.21-0.36 (4H, m), 0.69-0.89 (2H,m), 1.35 (9H, s), 3.64 (2H, dd, J=14.6 & 6.8 Hz), 4.14 (2H, dd, J=14.6 &6.8 Hz), 4.65 (1H, d, J=8.2 Hz), 6.45 (1H, d, J=8.2 Hz), 7.37-7.49 (2H,m), 7.63-7.77 (2H, m).

Elemental Analysis (for C₂₆ H₃₅ N₅ O₆)

Calcd.: C, 59.35; H, 6.97; N, 13.31

Found: C, 59.35; H, 6.84; N, 13.14.

Preparation 141,5-Bis-(cyclopropylmethyl)-3-(methoxyimino)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione15

Compound 15 is prepared in a manner similar to that described in Example1 below for preparation of Compound 8a in 87.1% yield.

M.p. =228°-230° C.

IR ν_(max) (KBr): 3443, 1714, 1678, 1660, 1598, 1503 cm⁻¹.

NMR (CDCl₃)δ: 0.93-1.08 (4H, m),1.15 (4H, m), 2.03 (2H, m), 3.96 (3H,s), 4.78 (2H, d, J=17.6 Hz), 4.90 (1H, d, J=17.6 Hz), 4.92 (2H, d,J=17.6 Hz), 7.17-7.33 (4H, m).

Elemental Analysis (for C₂₀ H₂₁ N₃ O₅)

Calcd.: C, 62.65; H, 5.52; N, 10.96

Found: C, 62.41; H, 5.62; N, 11.02.

Preparation 151,5-Bis-(cyclopropylcarbonylmethyl)-3-amino-1H-1,5-benzodiazepine-2,4(3H,5H)-dione16 h

1) To a solution of previously prepared1,5-bis-(cyclopropylcarbonylmethyl)-3-(t-butoxycarbonylamino)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione14 h (3.188 g) in ethyl acetate (16 ml) is added a solution of 4N HCl inethyl acetate (14 ml) under ice-cooling. After stirring the reactionmixture for 15 hr at room temperature, crystalline preparations arefiltered off. The resultant crystals are dissolved into methylenechloride/methanol (5:1) and washed with saturated aqueous sodiumhydrogencarbonate solution. The organic layer is dried over magnesiumsulfate and concentrated under reduced pressure. Crystallization from amixture of methylene chloride, methanol and diisopropyl ether yields thetitled Compound 16 h (2.092 g; yield, 84%).

M.p. =237°-238° C.

IR ν_(max) (KBr): 3375, 1700, 1667, 1600, 1505, 1416, 1389 cm⁻¹.

NMR (DMSO-d₆)δ: 0.79-1.03 (8H, m), 1.82-2.00 (2H, brs), 2.05-2.21 (2H,m), 4.08 (1H, s), 4.74(2H, d, J=18.2 Hz), 4.96 (2H, d, J=18.2 Hz),7.19-7.30 (2H, m), 7.30-7.40(2H, m).

Elemental Analysis (for C₁₉ H₂₁ N₃ O₄·0.3 H₂ O)

Calcd.: C, 63.25; H, 6.03; N, 11.65

Found: C, 63.35; H, 5.92; N, 11.65.

2) To a suspension of Compound 14 (3.34 g, 8.71 mmol) in methanol (320ml) is added 10% Pd/C (1.5 g) and the mixture is stirred for 20 hr underhydrogen gas. Organic substances are dissolved by the addition ofchloroform and the catalysts are filtered off. The residue obtained bydistilling the solvent under reduced pressure is dissolved intochloroform (100 ml) and methanol (300 ml), and again subjected to thereduction with 10% Pd/C catalyst for 20 hr. After removing the catalystsby filtration, the solvent is distilled under reduced pressure and theresidue is re-precipitated from methanol-ethyl acetate to obtainCompound 16 h (3.05 g, 98.5%).

Preparation 161,5-Bis-(pyrrolidinocarbonylmethyl)-3-amino-1H-1,5-benzodiazepine-2,4(3H,5H)-dione16a

Compound 16a is prepared by treating the previously obtained Compound14a in a manner similar to that described in Preparation 15 for thepreparation of Compound 16 h.

M.p. =246°-247° C.

IR ν_(max) (KBr): 3445, 1700, 1655, 1500, 1450, 1320 cm⁻¹.

NMR (DMSO-d₆)δ: 1.63-2.00 (10H, m), 3.23-3.38 (4H, m), 3.40-3.53 (4H,m), 4.04 (1H, s), 4.39(2H, d, J=16.6 Hz), 4.69 (2H, d, J=16.6 Hz),7.29-7.40 (2H, m), 7.40-7.51(2H, m).

Elemental Analysis (for C₂₁ H₂₇ N₅ O₄ ·0.7H₂ O)

Calcd.: C, 59.20; H, 6.72; N, 16.44

Found: C, 59.38; H, 6.46; N, 16.20.

Preparation 171,5-Bis-(thienylcarbonylmethyl)-3-amino-1H-1,5-benzodiazepine-2,4(3H,5H)-dione16g

Compound 16g is prepared by treating the previously obtained Compound14g in a manner similar to that described in Preparation 15, 1).

M.p. =208-209° C.

IR ν_(max) (KBr): 3450, 1700, 1679, 1662, 1585, 1502, 1419, 1241 cm⁻¹.

NMR (DMSO-d₆) δ: 1,96 (2H, s), 4.21 (1H, s), 5.18 (2H, d, J=18.0Hz),5.51 (2H, d, J=18.0Hz), 7.26-7.45 (6H, m), 8.08-8.20 (4H, m).

Elemental Analysis (for C₂₁ H₁₇ N₃ O₄ S₂)

Calcd.: C, 57.39; H, 3.90; N, 9.56; S, 14.59

Found: C, 57.28; H, 4.03; N, 9.40; S, 14.40.

Preparation 181,5-Bis-(cyclopropylmethyl)-3-amino-1H-1,5-benzodiazepine-2,4(3H,5H)-dione16o

Compound 16o is prepared by treating the previously obtained Compound14o in a manner similar to that described in Preparation 15, 1).

M.p. =147°-148° C.

IR ν_(max) (KBr): 3380, 1696, 1660, 1600, 1500, 1422 cm⁻¹.

NMR (DMSO-d₆)δ: 0.04-0.17 (4H, m), 0.22-0.35 (4H, m), 0.70-0.90 (2H, m),1.89 (2H, br.s), 3.63 (2H, dd, J=14.2 & 6.6 Hz), 3.89 (1H, s), 4.12 (2H,dd, J=14.2 & 6.6 Hz), 7.32-7.43 (2H, m), 7.58-7.71 (2H, m).

Elemental Analysis (for C₁₇ H₂₁ N₃ O₂)

Calcd.: C, 68.21; H, 7.07; N, 14.04

Found: C, 68.20; H, 7.12; N, 13.96.

Preparation 19 Ethyl 3-(N-BOC-amino)phenylthioacetate

To a solution of m-aminobenzenethiol (2.0 g, 15.9 mmol) in acetone (20ml) are added di-t-butyl dicarbonate (3.50 g, 16.04 mmol), then 5%aqueous sodium hydrogencarbonate solution (10 ml) under ice-cooling. Themixture is stirred at 0° C. for 30 min followed by at room temperaturefor 15 hr. After addition of ethyl acetate, the reaction mixture iswashed with water, dried over sodium sulfate and distilled under reducedpressure to remove the solvent. The resultant residue is purified bycolumn chromatography on silica gel (100 g silica gel; andhexane/toluene, 2:1) to obtain a product (3.09 g, 85.8%). To a solutionof the resultant compound (3.09 g) in acetone (50 ml) is added potassiumcarbonate (10 g). To the mixture is added dropwise ethyl bromoacetate(2.35 g) with stirring. After the reaction mixture is stirred for 3days, ether is added thereto and insoluble substances are removed byfiltration and the filtrate is distilled under reduced pressure. Theresultant residue is purified by column chromatography (100 g silicagel; and hexane/ethyl acetate, 3:1 to 2:1) and crystallization fromether/hexane to give the titled objective compound (2.4 g, 56.2%).

M.p. =70°-71° C.

IRν_(max) (KBr): 3424, 3353, 1717, 1599, 1538 cm⁻¹.

NMR (CDCl₃) δ: 1.24 (3H, t, J=7.3 Hz), 1.51 (9H, s), 3.64 (2H, s), 4.18(2H, q, J=7.3 Hz), 6.50 (1H, s), 7.00-7.11 (1H, m), 7.15-7.25 (2H, m),7.47 (1H, m).

Elemental Analysis (for C₁₅ H₂₈ NO₄ S)

Calcd.: C, 57.86; H, 6.80; N, 4.50; S, 10.30

Found: C, 57.61; H, 6.82; N, 4.67; S, 10.27.

Preparation 20 Ethyl 3-aminophenylthioacetate

A solution (4 ml) of 4N-hydrogen chloride in ethyl acetate is added to asolution of the above-mentioned BOC compound (1.255 g) in ethyl acetate(4 ml) and the mixture is stirred for 3 hr at room temperature. Afterthe addition of 5% sodium hydrogencarbonate (20 ml), the reactionmixture is extracted with ethyl acetate. The extract is washed withwater, dried over sodium sulfate and distilled to remove the solvent.The resultant crude amine (846 mg) is used in the next step withoutfurther purification.

NMR (CDCl₃)δ: 1.24 (3H, t, J=7.1 Hz), 3.62 (2H, s), 4.18 (2H, q, J=7.1Hz), 6.54 (1H, ddd, J=1.0, 2.3, 8.0 Hz), 6.72-6.81 (2H, m), 7.08 (1H, t,J=7.7 Hz). ##STR11##

EXAMPLE 11,5-Bis-(pyrrolidinocarbonylmethyl)-3-(N'-m-tolyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8a

The objective Compound 8a was prepared by N-alkylating a compoundprepared in Preparation 4, 5 or 6 in accordance with the generalprocedures described below.

Thus, a mixture of Compound 5, 6 or 7 (1.06 mmol) prepared inPreparation 4, 5 or 6 above and either of halides (2.40 mmol) preparedin Preparation 7 and 8, potassium carbonate (553 mg, 4.00 mmol) andpotassium iodide (80 mg, 0.48 mmol) in dimethylformamide is stirred for15 hr. The reaction mixture is poured into ice-cold water and extractedwith ethyl acetate. The extract is washed with water, dried over sodiumsulfate and distilled under reduced pressure to remove the solvent. Theresidue is purified by column chromatography on silica gel (100 g gel;and chloroform/methanol).

IR ν_(max) (KBr): 3421, 1694, 1655, 1557 cm⁻¹.

NMR (CDCl₃) δ: 1.70-2.05 (8H, m), 2.25 (3H, s), 3.47 (8H, m), 4.60 (4H,s), 5.23 (1H, m), 6.33 (1H, m), 6.79 (1H, m), 7.04-7.15 (3H, m), 7.22(1H, s), 7.28 (2H, m), 7.51 (2H, m).

Elemental Analysis (for C₂₉ H₃₄ N₆ O₅ ·0.6H₂ O)

Calcd: C, 62.41; H, 6.42; N, 15.04

Found: C, 62.49; H, 6.36; N, 15.08.

Compounds 8d, 8e, 8f, 8h, 8k, and 8l described in the following Exampleswere prepared in a manner similar to that described in Example 1 fromcorresponding starting materials.

EXAMPLE 21,5-Bis-(o-methylphenacyl)-3-{N'-(m-tolyl)-ureido}-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8d

M.p. =239°-242° C.

IR ν_(max) (KBr): 3408, 3363, 1691, 1670, 1643, 1614, 1600, 1571, 1501cm⁻¹.

NMR (CDCl₃ +CD₃ OD)δ: 2.29 (3H, s), 2.41 (6H, s), 5.07 (4H, s), 5.39(1H, s), 6.80 (1H, m), 7.14 (2H, m), 7.22-7.39 (9H, m), 7.45 (2H, m),7.65(2H, d, J=7.4 Hz).

Elemental Analysis (for C₃₅ H₃₂ N₄ O₅ ·0.5H₂ O)

Found: C, 70.28; H, 5.43; N, 9.52

Calcd.: C, 70.34; H, 5.57; N, 9.37.

EXAMPLE 31,5-Bis-(3,4-dimethoxyphenacyl)-3-(N'-(m-tolyl)-ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8e

Yield, quantitative; m.p. =266°-268° C.

IR ν_(max) (KBr): 3355, 1699, 1678, 1665, 1649, 1614, 1596, 1568 cm⁻¹.

NMR (DMSO+CD₃ OD)δ: 2.24 (3H,s), 3.85 (6H,s), 3.89 (6H,s), 5.15 (1H,d,(2 Hz), 5.29 (2H, d, J=18.0Hz), 5.55 (2H, d, J=18.0 Hz), 6.73 (1H, d,J=5.8 Hz), 6.89 (1H, d, J=8.2 Hz), 7.04-7.16 (4H, m), 7.20 (1H,s), 7.41(4H, s), 7.52 (2H, d, J=2.0 Hz), 7.74 (2H, dd, J=2.8, 8.2 Hz), 9.04 (1H,s).

Elemental Analysis (for C₃₇ H₃₆ N₄ O₉ ·0.4H₂ O)

Found: C, 64.60; H, 5.53; N, 8.10

Calcd.: C, 64.60; H, 5.39; N, 8.14.

EXAMPLE 41,5-Bis-(2-furylcarbonylmethyl)-3-{N'-(m-tolyl)-ureido}-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8f

M.p. =296°-298° C. (decomposition).

IR ν_(max) (nujol): 3367, 1706, 1689, 1663, 1644, 1570 cm⁻¹.

NMR (DMSO-d₆) δ: 2.22 (3H, s), 5.10 (1H, d, J=8.2 Hz), 5.12 (2H, d,J=18.0Hz), 5.38 (2H, d, J=18.0Hz), 6.73 (1H, s), 6.80 (2H, dd, J=3.6 &1.6 Hz), 7.10 (2H, d, J=4.4 Hz), 7.19 (1H, s), 7.37-7.55 (4H, m), 7.64(2H, d, J=3.6 Hz), 8.10 (2H, d, J=1.6 Hz), 9.05 (1H, s).

Elemental Analysis (for C₂₉ H₂₄ N₄ O₇ ·0.5H₂ O)

Found: C, 63.28; H, 4.57; N, 10.32

Calcd.: C, 63.38; H, 4.59; N, 10.20.

EXAMPLE 51,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-tolyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8 h

Yield, 74%; m.p. =246°-248° C.

IR ν_(max) (KBr): 3380, 1700, 1645, 1617, 1565, 1500, 1428

NMR (DMSO-d₆)δ: 0.79-1,06 (8H, m) 2.06-2.21 (2H, m), 2.22(3H, s), 4.82(2H, d, J=18.2 Hz), 4.99 (1H, d, J=8.0Hz), 4.99 (1H, d, J=18.2 Hz),6.68-6.79 (1H, m), 6.85 (1H, d, J-8.0Hz), 7.09 (1H, d, J=5.2 Hz), 7.18(1H, s), 7.28-7.50 (5H, m), 9.03 (1H, s).

Elemental Analysis (For C₂₇ H₂₈ N₄ O₅ ·0.1H₂ O)

Calcd.: C, 66.14; H, 5.80; N, 11.43

Found: C, 66.05; H, 5.80; N, 11.46.

EXAMPLE 61,5-Bis-(2-triphenylmethyltetrazol-5-ylmethyl)-3-(N'-(m-tolyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8k

Yield, 49.3%; m.p. =156°-158° C.

IR ν_(max) (KBr): 3411, 1711, 1678, 1612, 1597, 1546 cm⁻¹.

NMR (CDCl₃) δ: 2.27 (3H, s), 4.89 (2H, d, J=16.4 Hz), 5.05 (2H, d,J=16.4 Hz), 5.28 (1H, d, J=7.8 Hz), 6.24 (1H, d, J=7.8 Hz), 6.70-7.50(39H, m).

Elemental Analysis (for C₅₉ H₄₈ N₁₂ O·3H₂ O)

Found: C, 71.28; H, 5.15; N, 17.65

Calcd.: C, 71.50; H, 5.08; N, 16.96.

EXAMPLE 71,5-Bis-(tetrazol-5-ylmethyl)-3-(N'-(m-tolyl)-ureido)-1H-1,5-benzodiazepine-2,4-(3H,5H)-dione81

To a solution of Compound 8k (353 mg, 0.363 mmol) in methanol (20 ml) isadded 10% HCl (5 ml) and the mixture is stirred for 15 hr at roomtemperature. After the addition of water (150 ml), the mixture isextracted with ethyl acetate. The organic layer is washed with water andextracted with 10% aqueous sodium carbonate. The aqueous layer isacidified with 10% HCl and extracted with ethyl acetate. The extract iswashed with water, dried over sodium sulfate, and distilled underreduced pressure to remove the solvent. Crystallization frommethanol/benzene gives Compound 81 (118 mg, 66.7%).

M.p. =228-230° C.

IR ν_(max) (KBr): 3375, 3274, 1713, 1688, 1647, 1561 cm⁻¹.

NMR (CDCl₃)δ: 2.29 (3H, s), 5.22 (1H, s), 5.37 (2H, d, J=16.4 Hz), 5.41(2H, d, J=16.4 Hz), 6.83 (1H, m), 7.09-7.20 (3H, m), 7.45 (4H, m).

Elemental Analysis (for C₂₁ H₂ ON_(l2) O₃)

Found: C, 51.51; H, 4.23; N, 34.36

Calcd.: C, 51.64; H, 4.13; N, 34.41.

EXAMPLE 8 Disodium1,5-bis-(tetrazol-5-ylmethyl)-3-(N'-(m-tolyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8m

Disodium salt 8m is obtained quantitatively by lyophilizing a solutionof Compound 81 (85.8 mg, 0.176 mmol) in aqueous 0.1N NaOH solution (3.50ml).

IR ν_(max) (KBr): 3380, 1700, 1669, 1613, 1559 cm⁻¹.

NMR (DMSO-d₆)δ: 2.21 (3H, s), 4.83 (2H, d, J=15.6 Hz), 4.92 (1H d, J=8.2Hz), 5.24(2H, d, J=15.6 Hz), 6.70 (1H, d, J=5.0 Hz), 6.80 (1H, d, J=8.2Hz), 7.01-7.13 (2H,m), 7.18 (1H, s), 7.29 (2H, m), 7.88 (2H, m), 9.07(1H, s).

Elemental Analysis (for C₂₁ H₁₈ N₁₂ O₃ Na₂ ·1.6H₂ O)

Found: C, 44.80; H, 3.39; N, 30.09; Na, 8.26

Calcd.: C, 44.94; H, 3.81; N, 29.95; Na, 8.19.

Compounds 8n, 8o, 8p and 8q described in the following Examples wereprepared from corresponding starting materials in a manner similar tothat described in Example 1 above.

EXAMPLE 9 lb1,5-Bis-allyl-3-{N'-(m-tolyl)ureido}-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8n

M.p. =210°-211° C.

IR ν_(max) (nujol): 3327, 1702, 1666, 1641, 1561 cm⁻¹

NMR (DMSO-d₆)δ: 2.22 (3H, s), 4.45 (2H, dd, J=15.9 & 6.0 Hz), 4.67 (2H,dd, J=15.9 & 6.0 Hz), 4.90 (1H, d, J=7.6 Hz), 5.04-5.10 (2H, m), 5.14(2H, d, J=3.0Hz), 5.59-5.82 (2H, m), 6.68-6.78 (1H, m), 6.88 (1H, d,J=7.6 Hz), 7.08-7.14 (2H, m). 7.17 (1H, s), 7.35-7.48 (2H, m), 7.55-7.68(2H, m), 9.08 (1H, s).

Elemental Analysis (for C₂₃ H₂₄ N₄ O₃ ·0.3H₂ O)

Found: C, 67.50; H, 6.02; N, 13.65

Calcd.: C, 67.40; H, 6.05; N, 13.67.

EXAMPLE 101,5-Bis-(cyclopropylmethyl)-3-{N'-(m-tolyl)-ureido}-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8o

M.p. =221°-222° C.

IR ν_(max) (nujol): 3315, 1693, 1643, 1611 cm⁻¹.

NMR (DMSO-d₆)δ: 0.09-0.18 (4H, m), 0.23-0.37(4H, m), 0.72-0.92 (2H, m),2.22 (3H, s), 3.66 (2H, dd, J=14.2, 6.8 Hz), 4.16 (2H, dd, J=14.2, 6.8Hz), 4.79 (1H, d, J=7.6 Hz), 6.67-6.76 (1H, m), 6.84(1H, d, J=7.8 Hz),7.10 (2H, dd, J=3.0, 1.0 Hz), 7.18 (1H, s), 7.37-7.51 (2H, m), 7.66-7.78(2H, m), 9.09 (1H, s).

Elemental Analysis (for C₂₅ H₂₈ N₄ O₃ ·0.4H₂ O)

Found: C, 68.40; H, 6.50; N, 12.81

Calcd.: C, 68.29; H, 6.60; N, 12.74.

EXAMPLE 111,5-Bis-(o-methylbenzyl)-3-{N'-(m-tolyl)-ureido}-1H-l,5-benzodiazepine-2,4(3H,5H)-dione8p

M.p. =225°-227° C.

IR ν_(max) (nujol): 3334, 1701, 1688, 1641 cm⁻¹.

NMR (DMSO-d₆)δ: 2.12 (6H, s), 2.25 (3H, s), 4.99 (2H, d, J=16.4 Hz),5.09 (2H, d, J=16.4 Hz), 5.16 (1H, d, J=7.4 Hz), 6.75 (1H, d, J=6.2 Hz),6.88-7.23 (12H, m), 7.28-7.37 (2H, m), 7.45-7.56 (2H, m), 9.11 (1H, s).

Elemental Analysis (for C₃₃ H₃₂ N₄ O₃)

Found: C, 74.19; H, 6.15; N, 10.34

Calcd.: C, 74.41; H, 6.06; N, 10.52.

EXAMPLE 121,5-Bis-(m-BOC-aminophenylmethyl)-3-(N'-(m-tolyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8q

Yield, 65.3%.

IR ν_(max) (KBr): 3413, 1703, 1675, 1612, 1547 cm⁻¹.

NMR (CDCl₃)δ: 1.50 (18H, s), 2.10 (3H, s), 4.90 (4H, s), 5.47 (1H, d,J=8.2 Hz), 6.62 (2H, t, J=7.4 Hz), 6.70 (2H, d, J=8.0 Hz), 6.87 (1H, dJ=7.8 Hz), 6.87 (1H, d, J=7.8 Hz), 6.92 (1H, s), 7.04 (11H, m), 7.61(2H, d, J=8.2 Hz), 7.60 (1H, s).

Elemental Analysis (for C₄₁ H₄₆ N₆ O₇ ·0.5H₂ O)

Found: C, 66.31; H, 6.51; N, 11.08

Calcd.: C, 66.20; H, 5.37; N, 11.30.

EXAMPLE 131,5-Bis-(aminophenylmethyl)-3-(N'-(m-tolyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8r

To a solution of Compound 8q (360 mg, 0.490 mmol) in ethyl acetate (4ml) is added a solution (2 ml) of 4N HCl in ethyl acetate and themixture is stirred for 5 hr at room temperature. Crystals are filteredoff to obtain the objective compound (295 mg, 99.0%).

M.p.=176°-179° C.

IR ν_(max) (KBr): 3416, 2864, 2591, 1697, 1663, 1603, 1557 cm⁻¹.

NMR (CD₃ OD)δ: 2.29 (3H, s), 5.06 (2H, d, J=16.2 Hz), 5.25 (1H, s), 5.25(2H, d, J=16.2 Hz), 6.84 (1H, m), 7.11-7.18 (3H, m), 7.25-7.56 (12H, m).

Elemental Analysis (for C₃₁ H₃₂ N₆ O₃ C₁₂ ·H₂ O)

Found: C, 59.43; H, 5.69; N, 13.16; Cl, 11.54

Calcd.: C, 59.52; H, 5.48; N, 13.43; Cl, 11.33.

Compounds 8s and 8t described in the following Examples were preparedfrom corresponding starting materials in a manner similar to thatdescribed in Example 1 above.

EXAMPLE 141,5-Bis-(t-butoxycarbonylmethyl)-3-(N'-(m-tolyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8s

Yield 37.3%; m.p.=218°-219° C.

IR ν_(max) (KBr): 3370, 1742, 1708, 1653, 1615, 1570, 1500, 1422, 1370,1225, 1155 cm⁻¹.

NMR (CDCl₃ --CD₃ OD)δ: 1.44 (18H, s)2.28, (3H, s), 4.46, (4H, q, J=9.8Hz), 5.30 (1H, s), 6.16-6.30 (1H, br.s), 6.71-6.88 (2H, m), 7.03-7.23(3H, m). 7.36(4H, m).

Elemental Analysis (for C₂₉ H₃₆ N₄ O·2H₂ O)

Found: C, 62.59; H, 6.60; N, 10.10

Calcd.: C, 62.62; H, 6.60; N, 10.07.

NMR (CDCl₃ +CD₃ OD)δ: 2.28 (3H, s), 3.59 (2H, m), 3.76 (4H, m), 4.61(2H, m). 5.12 (1H, s), 6.80 (1H, m), 7.12 (2H, m), 7.21 (1H, s), 7.43(4H, m).

EXAMPLE 151,5-Bis-(2-hydroxymethyl)-3-(N'-(m-tolyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8t

M.p.=221°-224° C.

IR ν_(max) (KBr): 3437, 3387, 1701, 1629, 1561 cm⁻¹.

NMR (CDCl₃ +CD₃ OD)δ: 2.28 (3H, s), 3.59 (2H, m), 3.76 (4H, m), 4.61(2H, m), 5.12 (1H, s), 6.80 (1H, m), 7.12 (2H, m), 7.21 (1H, s), 7.43(4H, m).

Elemental Analysis (for C₂₁ H₂₄ N₄ O₃ ·0.2H₂ O)

Found: C, 60.53; H, 5.92; N, 13.55

Calcd.: C, 60.63; H, 5.91; N, 13.47.

EXAMPLE 161,5-Bis-(carboxymethyl)-3-(N'-(m-tolyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione8u

To a solution of Compound 8t (193 mg, 0.468 mmol) in acetone (24 ml) isadded dropwise Jones' reagent (3M, 1 ml) and mixture is stirred for 2 hrat 55° C. After cooling, water is added to the mixture. The mixture isextracted with chloroform/methanol (4:1) to obtain a crude carboxylicacid (166 mg, 80.6%).

NMR (CDCl₃ +CD₃ OD)δ: 2.27 (3H, s), 4.50 (2H, d, J=17.6 Hz), 4.55 (2H,d, J=17.6 Hz ), 5.30 (1H, s), 6.80 (1H, m), 7.11 (2H, m), 7.19 (1H, m),7.41 (4H, m).

EXAMPLE 17 1,5-Bis-(pyrrolidinecarbonylmethyl)-3-(N'-(3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 9a

Compound 6 is prepared in a manner similar to that described inPreparation 4 for the preparation of Compound 5 using Compound 4prepared in Preparation 3 and3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenyl-isocyanate. Compound9a is then prepared in a manner similar to that described in Preparation10 for the preparation of Compound 14 h using Compound 6 andpyrrolidinocarbonylmethyl bromide.

IR ν_(max) (CHCl₃)δ: 3433, 3372, 1708, 1656, 1599, 1558 cm⁻¹.

NMR (CDCl₃) δ: 1.73-2.02 (10H, m), 2.36 (2H, t, J=7.2 Hz), 3.19 (2H, m),3.30-3.53 (8H, m), 3.66 (3H, s), 4.57 (2H, d, J=16.8 Hz), 4.62 (2H, d,J=16.8 Hz), 4.96 (2H, s), 5.21 (1H, d, J=6.2 Hz), 5.49 (1H, br. s), 6.49(1H, d, J=7.6 Hz), 6.86 (1H, d, J=7.0 Hz), 7.12 (1H, d, J=7.8 Hz),7.23-7.40 (4H, m), 7.50 (2H, m), 7.98 (1H, s).

Elemental Analysis (for C₃₅ H₄₃ N₇ O₉ ·0.9H₂ O)

Found: C, 58.29; H, 6.15; N, 13.58

Calcd. : C, 58.23; H, 6.25; N, 13.58.

Compounds 9b, 9d, 9f, 9g, 9h, 9i, 9n, 9o, 9t, 9u, 10c, 10j, 10k, 10s,10t and 10u described in the following Examples were prepared in amanner similar to that described in Example 17 from correspondingstarting materials.

EXAMPLE 181,5-Bis-(thiazolidinecarbonylmethyl)-3-(N'-(3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione9b

IR ν_(max) (CHCl₃): 3431, 3374, 1709, 1663, 1598, 1558 cm⁻¹.

NMR (CDCl₃)δ: 1.82 (2H, qui, J=7.0 Hz), 2.35 (2H, t, J=7.3 Hz), 2.95(2H, t, J=6.1 Hz), 3.07 (2H, m), 3.19 (2H, m), 3.66 (3H, s), 3.69 (2H,m), 3.82 (2H, t, J=6.2 Hz), 4.38-4.70 (8H, m), 4.97 (2H, s), 5.24 (1H,br.s), 5.42 (1H, br.s), 6.55 (1H, br.s), 6.89 (1H, d, J=7.8 Hz), 7.14(1H, d, J=7.8 Hz), 7.23-7.52 (7H, m), 7.88 (1H, s).

Elemental Analysis (for C₃₃ H₃₉ N₇ O₉ S₂)

Found: C, 52.68; H, 5.43; N, 12.91; S, 7.83

Calcd.: C, 52.66; H, 5.38; N, 13.03; S, 8.52.

EXAMPLE 191,5-Bis-(o-methylphenacyl)-3-(N'-(3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione9d

M.p.=162°-164° C.

IR ν_(max) (nujol): 3353, 1703, 1689, 1527 cm⁻¹.

NMR (DMSO-d₆)δ: 1.81 (2H, quintet, J=7.4 Hz), 2.34 (2H, t, J=7.4 Hz),2.35 (6H, s), 3.18 (2H, q, J=6.6 Hz), 3.65 (3H, s), 4.91 (2H, s), 4.97(4H, s), 5.25-5.36 (1H, br.s), 5.40 (1H, d, J=6.6 Hz), 6.66-6.77 (1H,br.s), 6.79-6.88 (1H, m), 7.00-7.65 (16H, m).

Elemental Analysis (for C₄₁ H₄₁ N₅ O₉)

Found: C, 65.59; H, 5.58; N, 9.34

Calcd.: C, 65.85; H, 5.53; N, 9.37.

EXAMPLE 201,5-Bis-(2-furylcarbonylmethyl)-3-(N'-(3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 9f

M.p.=176°-178° C.

IR ν_(max) (nujol): 3346, 3122, 1727, 1706, 1685, 1570 cm⁻¹.

NMR (CDCl₃)δ: 1.81 (2H, quintet, J=7.0 Hz), 2.34 (2H, t, J=7.4 Hz), 3.18(2H, q, J=6.0 Hz), 3.65 (3H, s), 4.98 (2H, s), 5.11 (2H, d, J=17.8 Hz),5.28 (2H, d, J=17.8 Hz), 5.44 (1H, d, J=7.0 Hz), 6.56 (2H, d.d, J=1.8 &0.6 Hz), 6.90 (1H, d, J=7.2 Hz), 7.09-7.43 (11H, m), 7.61 (2H, d.d,J=0.6 & 0.6 Hz).

Elemental Analysis (for C₃₅ H₃₃ N₁₁ ·0.5H₂ O)

Found: C, 59.33; H, 4.75; N, 9.75

Calcd.: C, 59.32; H, 4.84; N, 9.88.

EXAMPLE 211,5-Bis-(2-thienylcarbonylmethyl)-3-(N'-(3-(3-(carbomethoxy)propylcarbamoylmethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione9g

M.p.=197°-198° C.

IR ν_(max) (nujol): 3338, 1705, 1679, 1667, 1637, 1568 cm⁻¹.

NMR (DMSO-d₆)δ: 1.64 (2H, qui, J=7.0Hz), 2.30(2H, t, J=7.4 Hz), 3.00 (2HJ=5.8 Hz), 3.57 (3H, s), 4.93 (2H, s), 5.12 (1H, d, J=7.8 Hz), 5.28 (2H,d, J=18.0 Hz), 5.56 (2H, J=18.0Hz), 6.84-6.96 (2H, m), 7.12-7.55 (1OH,m) 8.08-8.22 (4H, m), 9.18 (1H, s).

Elemental Analysis (for C₃₅ H₃₃ N₅ O₉ S₂ ·0.5H₂ O)

Found: C, 56.60; H, 4.54; N, 9.34; S, 8.91

Calcd.: C, 56.75; H, 4.63; N, 9.45; S, 8.66.

EXAMPLE 221,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione9h

IR ν_(max) (KBr): 3387, 3367, 1711, 1691, 1615, 1598, 1556, 1502 cm⁻¹.

NMR (DMSO-d₆)δ: 0.80-1.08 (8H, m), 1.64 (2H, qui, J=7.1 Hz), 2.15 (2H,m), 2.30 (2H, t, J=7.4 Hz), 2.99 (2H, q, J=6.2 Hz), 3.57 (3H, s), 4.84(2H, d, J=18.0 Hz),4.92 (2H, s), 4.99 (2H, d, J=18.0 Hz), 4.99 (1H, d,J=8.0Hz), 6.88 (2H, m), 7.15-7.47 (8H, m), 9.16 (1H, s).

Elemental Analysis (for C₃₃ H₃₇ N₅ O₉ ·H₂ O)

Found: C, 59.62; H, 5.74; N, 10.22

Calcd.: C, 59.54; H, 5.90; N, 10.52.

EXAMPLE 231,5-Bis-(cyclopentylcarbonylmethyl)-3-(N'-(3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione9i

IR ν_(max) (KBr): 3382, 1723, 1702, 1614, 1598, 1557 cm⁻¹.

NMR (CDCl₃)δ: 1.45-2.00 (18H, m), 2.36 (2H, t, J=7.2 Hz), 2.94 (2H, qui,J=7.9 Hz), 3.20 (2H, q, J=6.5 Hz), 3.67 (3H, s), 4.68 (2H, d, J=18.0Hz),4.79 (2H, d, J=18.0Hz), 4.97 (2H, s), 5.24 (1H, br.s), 5.30 (1H, d,J=7.4 Hz), 6.45 (1H, d, J=7.4 Hz), 6.90 (1H, d, J=7.4 Hz), 7.09-7.35(7H, m), 7.39 (1H, s).

Elemental Analysis (for C₃₇ H₄₅ N₅ O₉ ·0.5H₂ O)

Found: C, 62.26; H, 6.43; N, 10.04

Calcd.: C, 62.35; H, 6.50; N, 9.83.

EXAMPLE 241,5-Bis-allyl-3-(N'-(3-(3-(carbomethoxy)-propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione9n

M.p.=196°-197° C.

IR ν_(max) (nujol): 3423, 3288, 1729, 1689, 1638 cm⁻¹.

NMR (DMSO-d₆)δ: 1.83 (2H, quintet, J=6.8 Hz), 2.36 (2H, t, J=7.4 Hz),3.21 (2H, q, J=7.0Hz), 3.67 (3H, s), 4.54 (4H, brs), 4.95 (2H, s), 5.16(2H, s), 5.23 (1H, d, J=6.0 Hz), 5.25 (2H, s), 5.73-5.96 (2H, m),6.64-6.76 (1H, m), 6.90 (1H, d, J=6.6 Hz), 7.05-7.36 (6H, m), 7.40-7.58(3H, m).

Elemental Analysis (for C₂₉ H₃₃ N₅ O₇)

Found: C, 61.57; H, 5.89; N, 12.43

Calcd.: C, 61.80; H, 5.90; N, 12.43.

EXAMPLE 251,5-Bis-(cyclopropylmethyl)-3-(N'-(3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione9o

M.p.=215°-216° C.

IR ν_(max) (nujol): 3421, 2184, 1729, 1685, 1637 cm⁻¹.

NMR (DMSO-d₆) 0.09-0.18 (4H,m), 1.64 (2, quintet, J=7.0Hz), 2.30 (2H, t,J=7.4 Hz), 2.99 (2H, q, J=6.6 Hz), 3.57 (3H, s), 3.66 (2H, d.d, J=14.0 &7.0 Hz), 4.16 (2H, d.d, J=14.0 & 7.0Hz), 4.79 (1H, d, J=7.4 Hz), 4.91(2H, s), 6.77-6.91 (2H, m), 7.13-7.32 (3H, m), 7.35, (1H, s), 7.38-7.49(2H, m), 7.67-7.78 (2H, m), 9.21 (1H, s).

Elemental Analysis (for C₃₁ H₃₇ N₅ O₇ ·0.3H₂ O)

Found: C, 62.64; H, 6.28; N, 11.76

Calcd.: C, 62.36; H, 6.35; N, 11.73.

EXAMPLE 261,5-Bis-(2-hydroxyethyl)-3-(N'-(3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione9t

M.p.=147°-149° C.

IR ν_(max) (KBr): 3412, 3308, 1725, 1691, 1664, 1642, 1599, 1565 cm⁻¹.

NMR (CDCl₃ +CD₃ OD)δ: 1.81 (2H, qui, J=7.1 Hz), 2.36 (2H, t, J=7.4 Hz),3.18 (2H, t, J=6.9 Hz), 3.48-3.83 (6H, m), 3.66 (3H, s), 4.63 (2H, m),4.97 (2H, s), 5.11 (1H, s), 6.91 (1H, d, J=7.2 Hz), 7.12-7.53 (7H, m).

Elemental Analysis (for C₂₇ H₃₃ N₅ O₉ ·0.3H₂ O)

Found: C, 56.15; H, 5.92; N, 12.32

Calcd.: C, 56.21; H, 5.87; N, 12.14.

EXAMPLE 271,5-Bis-(carboxymethyl)-3-(N'-(3-(3-(carbomethoxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione9u

NMR (CDCl₃ +CD₃ OD)δ: 1.81 (2H, qui, J=7.0 Hz), 2.36 (2H, t, J=7.4Hz),3.18 (2H, t, J=7.1 Hz), 3.66 (3H, s), 4.49 (2H, d, J=17.6 Hz), 4.56(2H,d, J=17.6 Hz), 5.00 (2H, s), 5.30 (1H, s), 6.93 (1H, d, J=7.4 Hz),7.13-7.52 (7H, m).

EXAMPLE 281,5-Bis-(cyclopropylcarbamoylmethyl)-3-(N'-(3-(3-(benzoyloxycarbonyl)propylcarbamoyloxymethyl)-phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 10c

IR ν_(max) (CHCl₃): 3333, 1710, 1668, 1614, 1599, 1560, 1521 cm⁻¹.

NMR (CDCl₃ +CD₃ OD)δ: 0.44 (4H, m), 0.67 (4H, m), 1.84 (2H, qui, J=7.1Hz), 2.41 (2H, t, J=7.5 Hz), 2.63 (2H, m), 3.19 (2H, t, J=6.8 Hz),4.34(2H, d, J=16.2 Hz), 4.70 (2H, d, J=16.2 Hz), 5.00 (2H, s), 5.11 (2H,s), 5.13 (1H, s), 6.93 (1H, d, J=7.2 Hz), 7.14-7.52 (12H, m).

Elemental Analysis (for C₃₉ H₄₃ N₇ O₉ ·0.6H₂ O)

Found: C, 61.28; H, 5.75; N, 12.83

Calcd.: C, 61.62; H, 5.83; N, 12.82.

EXAMPLE 291,5-Bis-(1-methylimidazol-2-ylmethyl)-3-(N'-(3-(3-(benzoyloxycarbonyl)propylcarbamoyloxymethyl)-phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione10j

Compound 10j is prepared using 1-methyl-2-chloromethylimidazole (T. S.Manoharan and R. S. Brown, J. Org. Chem., 54, 1439 (1989)) as a startingmaterial in 84.9 % yield.

NMR (CDCl₃)δ: 1.81 (2H, qui, J=7.0Hz), 2.38 (2H, t, J=7.5 Hz), 3.16 (2H,t, J=6.6 Hz), 3.53 (6H, s), 4.91-5.19 (5H, m), 4.94 (2H, s), 5.10 (2H,s), 5.44 (1H, t, J=4.6 Hz), 6.74 (3H, s), 6.86 (2H, s), 6.89 (1H, s),7.11 (1H, t, J=7.6 Hz), 7.18-7.37 (5H, m), 7.33 (5H, s), 7.76 (2H, m),8.10 (1H, br.s).

Elemental Analysis (for C₃₉ H₄₁ N₉ O₇ ·0.6H₂ O)

Found: C, 61.77; H, 5.69; N, 16.76

Calcd.: C, 61.75; H, 5.61; N, 16.62.

EXAMPLE 301,5-Bis-(2-triphenylmethyltetrazol-5-yl)-3-(N'-(3-(3-(benzyloxycarbonyl)propylcarbamoyloxymethyl)-phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione10k

Yield, 75.5%.

IR ν_(max) (KBr): 3405, 1714, 1639, 1614, 1599, 1558 cm⁻¹.

NMR (CDCl₃)δ: 1.81 (2H, qui, J=7.2 Hz), 2.37 (2H, t, J=7.3 Hz), 3.16(2H, q, J=6.6 Hz), 4.88 (2H, d, J=16.4 Hz), 4.96 (2H, s), 5.06 (2H, d,J=16.4 Hz), 5.09 (2H, s), 5.16 (1H, m), 5.28 (1H, d, J=7.6 Hz), 6.45(1H, d, J=7.6 Hz), 6.89-7.02 (12H, m), 7.08-7.38 (30H, m), 7.45 (2H, m).

Elemental Analysis (for C₇₁ H₆₁ N₁₃ O₇ ·0.7H₂ O)

Found: C, 69.88; H, 5.34; N, 14.96

Calcd.: C, 69.85; H, 5.15; N, 14.91.

EXAMPLE 311,5-Bis-(tert-butoxycarbonylmethyl)-3-(N'-(3-(3-(benzyloxycarbonyl)propylcarbamoyloxymethyl)phenyl)-ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione10s

Yield, 94.3%.

IR ν_(max) (KBr): 3385, 1739, 1711, 16147, 1598, 1558 cm⁻¹.

NMR (CDCl₃) δ: 1.44 (18H, s), 1.84 (2H, qui, J=7.1 Hz), 2.41 (2H, t,J=7.5 Hz), 3.20 (2H, q, J=6.0 Hz), 4.37 (2H, d, J=17.2 Hz), 4.56 (2H, d,J=17.2 Hz), 4.93 (2H, s), 5.11 (2H, s), 5.28 (1H, br.s), 5.33 (1H, d,J=7.4 Hz),6.60 (1H, d, J=8.2 Hz), 6.86 (1H, d, J=7.4 Hz), 7.05-7.40(12H, m), 7.43 (1H, s).

Elemental Analysis (for C₄₁ H₄₉ N₅ O₁₁ ·0.4H₂ O)

Found: C, 61.97; H, 6.33; N, 8.86

Calcd.: C, 61.94; H, 6.31; N, 8.81.

EXAMPLE 321,5-Bis-(2-hydroxyethyl)-3-(N'-(3-(3-(benzyloxycarbonyl)propylcarbamoyloxymethyl)phenyl)-ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione10t

M.p.=138°-142° C.

IR ν_(max) (CHCl₃): 3449, 3375, 1703, 1675, 1658, 1599, 1556 cm⁻¹.

NMR (CDCl₃ +CD₃ OD)δ: 1.83 (2H, qui, J=7.2 Hz), 2.41 (2H, t, J=7.4 Hz),3.18 (2H, t, J=6.9 Hz), 3.51-3.66 (2H, m), 3.67-3.84 (4H, m), 4.52-4.70(2H, m), 4.97 (2H, s), 5.11 (3H, s), 6.91 (1H, d, J=7.2 Hz), 7.17(1H, t,J=7.4 Hz), 7.22-7.42 (10H, m), 7.47 (2H, m).

Elemental Analysis (for C₃₃ H₃₇ N₅ O₉ ·0.5H₂ O)

Found: C, 60.35; H, 5.97; N, 10.83

Calcd.: C, 60.36; H, 5.83; N, 10.66.

EXAMPLE 331,5-Bis-(carboxymethyl)-3-(N'-(3-(3-(benzyloxycarbonyl)propylcarbamoyloxymethyl)-phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione10u

NMR (CDCl₃ +CD₃ OD)δ: 1.84 (2H, qui, J=7.1 Hz), 2.41(2H, t, J=7.2 Hz),3.1-3.3 (2H, m), 4.49 (2H, d, J=18.0Hz), 4.56 (2H, d, J=18.0Hz), 5.01(2H, s), 5.11 (1H, s), 5.30 (1H, s), 6.93 (1H, d, J=7.2 Hz), 7.14-7.50(12H, m).

EXAMPLE 341,5-Bis-(pyrrolidinecarbonylmethyl)-3-(N'-(3-(3-(carboxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione11a

To a solution of Compound 9a (243 mg, 0.344 mmol) in methanol (3.5 ml)is added a solution of lithium hydroxide monohydrate (35 mg, 0.834 mmol)in water (0.6 ml) and the mixture is stirred at room temperature for 7hr. After the addition of water, the mixture is washed with chloroform.After acidifying with 10% HCl, the mixture is extracted with chloroformcontaining 10% methanol. The extract is washed with water, dried oversodium sulfate, and distilled under reduced pressure to remove thesolvent to obtain Compound 11a (224 mg, 94.1%).

NMR (CDCl₃ +CD₃ ODδ: 1.73-2.02 (10H, m), 2.34 (2H, t, J=7.3 Hz), 3.21(2H, t, J=6.6 Hz), 3.40-3.56 (8H, m), 4.62 (4H, s), 5.03 (2H, s), 5.26(1H, s), 6.90 (1H, d, J=7.0 Hz), 7.14-7.55 (7H, m).

Compounds 11 described in the following Examples were prepared in amanner similar to that described in Example 34 above.

EXAMPLE 351,5-Bis-(cyclopropylcarbamoylmethyl)-3-(N'-(3-(3-(carboxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione 11c

NMR (CDCl₃ +CD₃ OD)δ: 0.44 (4H, m), 0.67 (4H, m), 1.81 (2H, m), 2.34(2H, t, J=7.2 Hz), 2.63 (2H, m), 3.1-3.3 (2H, m), 4.34 (2H, d, J=16.0Hz), 4.70 (2H, d, J=16.0 Hz), 5.01 (2H, s), 5.13 (1H, s), 6.92 (1H, d,J=7.4 Hz), 7.14-7.46 (7H, m).

EXAMPLE 361,5-Bis-(o-methylphenacyl)-3-(N'-(3-(3-(carboxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione11d

M.p.=174°-176° C.

IR ν_(max) (nujol): 3351, 3312, 1702, 1689, 1668, 1645, 1618, 1572 cm⁻¹.

NMR (DMSO-d₆)δ: 1.61 (2H, quintet, J=7.0 Hz), 2.20 (2H, t, J=7.4 Hz),2.36 (6H, s), 2.99 (2H, q, J=6.8 Hz), 4.93 (2H, s), 5.07(1H, d, J=8.0Hz), 5.09 (2H, d, J=17.6 Hz), 5.20 (2H, d, J=17.6 Hz), 6.84-6.96 (2H,m), 7.16-7.41 (7H, m), 7.44-7.56 (6H, m), 7.75-7.83 (2H, m), 9.16(1H,s).

Elemental Analysis (for C₄₀ H₃₉ N₅ O₉ ·0.3H₂ O)

Found : C,64.33; H, 5.28; N, 9.31

Calcd.: C,61.80; H, 6.15; N, 12.01.

EXAMPLE 371,5-Bis-(2-furylcarbonylmethyl)-3-(N'-(3-(3-(carboxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione11f

M.p.=157°-160° C.

IR ν_(max) (nujol): 3507, 3439, 3345, 3127, 1707, 1679, 1664, 1651,1618, 1571 cm⁻¹.

NMR (DMSO-d₆)δ: 1.61 (2H, quintet, J=7.0 Hz), 2.21 (2H, t, J=7.2 Hz),2.99 (2H, q, J=8.0Hz), 3.34 (2H, s), 4.92 (2H, s), 5.10 (1H, d,J=8.0Hz), 5.12 (2H, d, J=18.2 Hz), 5.38 (2H, d, J=18.2 Hz), 6.80 (2H,d.d, J=3.6 & 1.8 Hz), 6.87 (1H, s), 6.90 (1H, s), 7.13-7.54 (8H, m),7.65 (2H, d.d, J=3.6 & 0.6 Hz), 8.10 (2H, d.d, J=1.8 & 0.6 Hz), 9.18(1H, s).

Elemental Analysis (for C₃₄ H₃₁ N₅ O₁₁ ·H₂ O)

Found: C, 58.16; H, 4.68; N, 9.86

Calcd.: C, 58.04; H, 4.73; N, 9.95.

EXAMPLE 381,5-Bis-(2-thienylcarbonylmethyl)-3-(N'-(3-(3-(carboxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione11g

M.p.=204°-207° C.

IR ν_(max) (KBr): 3340, 1705, 1666, 1637, 1614, 1568 cm⁻¹.

NMR (DMSO-d₆)δ: 1.61 (2H, quintet, J=7.0Hz), 2.21 (2H, t, J=7.4 Hz),2.99 (2H, q, J=5.8 Hz), 4.92 (2H, s), 5.12 (1H, s, J=8.0Hz), 5.29 (2H,d, J=18.0Hz), 5.56 (2H, d, J=18.0Hz), 6.81-6.97 (2H, m), 7.12-7.55 (1OH,m), 8.09-8.21 (4H, m), 9.19 (1H, s).

Elemental Analysis (for C₃₄ H₃₁ N₅ O₉ S₂ ·0.5H₂ O)

Found : C, 56.40; H, 4.35; N, 9.49; S, 9.11

Calcd.: C, 56.19; H, 4.44; N, 9.64; S, 8.82.

EXAMPLE 391,5-Bis-allyl-3-(N'-(3-(3-(carboxy)propyl-carbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione11n

M.p.=108°-112° C.

IR ν_(max) (nujol): 3311, 1695, 1664, 1640, 1616, 1598, 1563 cm⁻¹.

NMR (DMSO-d₆)δ: 1.61 (2H, quintet, J=7.4 Hz), 2.20 (2H, t, J=7.4 Hz),2.99 (2H, q, J=6.4 Hz), 4.45 (2H, d.d, J=16.0 & 5.8 Hz), 4.67 (2H, d.d,J=16.0 & 5.80Hz), 4.90 (1H, d, J=7.6 Hz), 4.92 (2H, s), 5.08 (2H, s),5.15 (2H, d, J=3.2 Hz), 5.59-5.82 (2H, m), 6.83-6.95 (2H, m), 7.14-7.32(3H, m), 7.34 (1H, s), 7.35-7.48 (2H, m), 7.55-7.68 (2H, m), 9.22 (1H,s).

Elemental Analysis (for C₂₈ H₃₁ N₅ O₇ ·1.5H₂ O)

Found : C, 60.02; H, 5.73; N, 12.60

Calcd.: C, 60.21; H, 5.77; N, 12.54.

EXAMPLE 401,5-Bis-(cyclopropylmethyl)-3-(N'-(3-(3-(carboxy)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione11o

M.p.=147°-149° C.

NMR (DMSO-d₆)δ: 0.04-0.17 (4H, m), 0.28-0.35 (4H, m), 0.72-0.92 (2H, m),1.61 (2H, quintet, J=7.0Hz), 2.30 (2H, t, J=7.4 Hz), 2.99 (2H, q, J=6.6Hz), 3.66 (2H, d.d, J=14.0 & 7.0Hz), 4.16 (2H, d.d, J=14.0 & 7.0 Hz),4.79 (1H, d, J=7.4 Hz), 4.91 (2H, s), 6.77-6.91 (2H, m), 7.13-7.32 (3H,m), 7.35 (1H, s), 7.38-7.49 (2H, m), 7.67-7.78 (2H, m), 5 9.21 (1H, s).

Elemental Analysis (for C₃₀ H₃₅ N₅ O₇ S₂ ·0.3H₂ O)

Found : C, 61.81; H, 6.11; N, 11.97

Calcd.: C, 61.80; H, 6.15; N, 12.01. ##STR12##

EXAMPLE 411,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-carboethoxyphenyl))ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-i

To a solution of methyl 3-aminobenzoate (200 mg, 1.21 mmol) intetrahydrofuran (10 ml) are added successively triphosgene (126 mg,0.424 mmol) and triethylamine (354μ, 2.54 mmol). After stirring for 1510 min, previously prepared amine 16 h (300 mg, 0.844 mmol) is added andthe stirring is continued for another 3 hr. The reaction mixture ispartitioned between a mixture of methylene chloride/methanol (5:1) andwater. The organic layer is washed with water, dried over magnesiumsulfate and concentrated under reduced pressure. The resultant crudeproduct is purified by column chromatography on silica gel(toluene/ethyl acetate, 1:1) and recrystallized from a mixture ofmethylene chloride, methanol and diisopropyl ether to give the titledCompound 17i (320 mg; yield, 69%).

M.p.=222°-223° C.

IR ν_(max) (KBr): 3400, 1708, 1599, 1562, 1503, 1425 cm⁻¹.

NMR (DMSO-d₆)δ: 0.79-1.04 (8H, m), 1.29 (3H, t, J=7.0 Hz), 2.06-2.22(2H, m), 4.28 (2H, q, J=7.0Hz), 4.83 (2H, d, J=18.0Hz), 5.00 (1H, d,J=7.8 Hz), 5.00 (2H, d, J=18.0 Hz), 6.89 (1H, d, J=7.8 Hz), 7.27-7.60(7H, m), 8.60 (1H, s), 9.39 (1H, s).

Elemental Analysis (for C₂₉ H₃₀ N₄ O₇ ·0.3H₂ O)

Calcd.: C, 63.10; H, 5.59; N, 10.15

Found: C, 63.05; H, 5.61; N, 10.28.

EXAMPLE 421,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-carboxyphenyl))ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-ii

Compound 17-ii is prepared by hydrolyzing the ester 17i obtained inExample 41 with an alkali in a conventional manner.

M.p.=274°-276° C.

IR ν_(max) (KBr): 3405, 1702, 1561, 1500, 1431 cm⁻¹.

NMR (DMSO-d6)δ: 0.75-1.05 (8H, m), 2.06-2.24 (2H, m), 4.83 (2H, d,J=18.2 Hz), 5.00 (1H, d, J=7.8 Hz), 5.00 (2H, d, J=18.2 Hz), 6.88 (1H,d, J=7.8 Hz), 7.27-7.55 (7H, m), 8.00 (1H, s), 9.30 (1H, s).

Elemental Analysis (for C₂₇ H₂₆ N₄ O₇ ·0.5H₂ O)

Calcd.: C, 61.47; H, 5.16; N, 10.62

Found: C, 61.47; H, 5,15; N, 10.62.

EXAMPLE 431,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(isopropylsulfonylaminocarbonyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-iii

Compound 17-iii is prepared in a manner similar to that described aboveusing amine 16 h and 3-isopropylsulfonylaniline (disclosed in EP 0 508796 A1) as starting materials.

M.p.=193°-194° C.

IR ν_(max) (KBr): 3380, 1700, 1602, 1553, 1503, 1432 cm⁻¹.

NMR (DMSO-d₆)δ: 0.80-1.05 (8H, m), 1.22 (6H, d, J=6.8 Hz), 2.06-2.23(2H, m), 3.55-3.76 (1H, broad), 4.83 (2H, d, J=18.2 Hz), 5.00 (1H, d,J=8.0Hz), 5.00 (2H, d, J=18.2 Hz), 6.87 (1H, d, J=8.0 Hz), 7.21-7,56(7H, m), 7.85 (1H, s), 9.33 (1H, s).

Elemental Analysis (for C₃₀ H₃₃ N₅ O₈ S·2.2H₂ O)

Calcd.: C, 54.32; H, 5.68; N, 10.56; S, 4.83

Found: C, 54.16; H, 5.58; N, 10.69; S, 4.88.

EXAMPLE 441,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(tetrazoyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-iv

To a suspension of 3-amino-(1H-tetrazol-5-yl)benzene (disclosed in EP 0508 796 A1) (195 mg, 1.21 mmol.) in tetrahydrofuran (10 ml) is addedtriethylamine (177 μl, 1.27 mmol) under ice-cooling and the mixture isstirred for 5 min. After addition of triphosgene (126 mg, 0.42 mmol) andtriethylamine (354 μl, 2.54 mmol) in series, the reaction mixture isstirred for 15 min. To the mixture is added previously prepared amine 16h (300 mg, 0.844 mmol) and the mixture is stirred at 0° C.for 30 min,then at room temperature for 3 hr. The reaction mixture is concentratedand partitioned between a mixture of methylene chloride-methanol (5:1)and water. The organic layer is dried over magnesium sulfate andconcentrated under reduced pressure. The resultant crude product ispurified by chromatography on silica gel (methylene chloride/methanol(5:1)) and recrystallized from a mixture of methanol and ether to givethe titled Compound 17-iv (220 mg; yield, 48%) as white crystals.

M.p.=213°-215° C.

IR ν_(max) (KBr): 3400, 1685, 1660, 1600, 1550, 1502, 1433 cm⁻¹.

NMR (DMSO-d₆)δ: 0.72-1.04 (8H, m), 2.06-2.25 (2H, m), 4.84 (2H, d,J=18.4 Hz), 5.01 (2H, d, J=18.4 Hz), 5.03 (1H, d, J=7.8 Hz), 6.93 (1H,d, J=7.8 Hz), 7.26-7,60 (8H, m), 8.04 (1H, s), 9.31 (1H, s).

Elemental Analysis (for C₂₇ H₂₆ N₈ O₅ ·1.5H₂ O)

Calcd.: C, 56.94; H, 5.13; N, 19.67

Found: C, 56.72; H, 5.04; N, 19.38.

EXAMPLE 451,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(carboethoxymethoxy)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-v

1) Ethyl 3-(N-tert-butyloxycarbonylamino)phenyloxyacetate

To a solution of 3-aminophenol (2.18 g, 20 mmol) in tetrahydrofuran (10ml) is added di-tert-butyl dicarbonate (4.578 g, 21 mmol) and themixture is stirred for 15 hr at room temperature. To the residueobtained by concentrating the reaction mixture is purified bychromatography on silica gel (hexane/ethyl acetate (5:1)). To a solutionof the resultant oily residue of 3-(N-tert-butyloxycarbonylamino)phenolin dimethylformamide are added ethyl 2-bromoacetate (2.44 ml, 22 mmol),potassium iodide (183 mg, 1.1 mmol) and potassium carbonate (3.04 g, 22mmol) and the mixture is stirred for 15 hr at room temperature. Thereaction mixture is concentrated under reduced pressure and theconcentrate is partitioned between a mixture of ethyl acetate and water.The organic layer is dried over magnesium sulfate and concentrated underreduced pressure. The resultant crude product is purified bychromatography on silica gel (toluene/ethyl acetate (9:1)) to give thetitled compound as colorless oil.

NMR (CDCl₃)δ: 1.22 (3H, t, J=7.4 Hz), 1.47 (9H, s), 4.17 (2H, q, J=7.4Hz), 4.70 (2H, s), 6.47-6.57 (1H, m), 6.99-7.21(3H, m), 9.34 (1H, s).

2) Ethyl 3-aminophenyloxyacetate

To a solution of the compound prepared in 1) above in ethyl acetate (38ml) is added 4N HCl (solution in ethyl acetate) (25 ml) and the mixtureis stirred for 3 hr at room temperature. The crystalline precipitatesare filtered off. The resultant crystals are dissolved in methylenechloride and washed with saturated aqueous sodium hydrogencarbonatesolution. The organic layer is dried over magnesium sulfate andconcentrated under reduced pressure to obtain the titled compound (1.744g; yield from 3-aminophenol, 45%) as colorless oil.

NMR(CDCl₃) δ: 1.30 (3H, t, J=7.4 Hz), 4.27 (2H, q, J=7.4 Hz), 4.57 (2H,s), 6.23-6.38 (3H, m), 7.00-7.11 (1H,m).

The titled Compound 17-v is prepared in a manner similar to thatdescribed in Example 41 for the preparation of Compound 17i using3-aminophenoxyacetic acid prepared in (2) above and the previouslyprepared amine 16 h.

M.p.=205°-206° C.

IR ν_(max) (KBr): 3370, 1760, 1700, 1642, 1608, 1602, 1562, 1500, 1455,1428 cm⁻¹.

NMR (DMSO-d₆)δ: 0.77-1.08 (8H, m), 1.19 (3H, t, J=7.0Hz), 2.06-2.23 (2H,m), 4.15 (2H, q, J=7.0 Hz), 4.86 (2H, s), 4.83 (2H, d, J=18.0 Hz), 4.99(1H, d, J=8.0Hz), 5.00 (2H, d, J=18.0Hz), 6.46 (1H, d.d, J=8.2 & 2.4Hz), 6.79-6.92 (2H, m), 6.84 1H, s), 7.12 (1H, t, J=8.2 z), 7.27-7,50(4H, m), 9.16 (1H, s).

Elemental Analysis (for C₃₀ H₃₂ N₄ O₈ ·0.3H₂ O)

Calcd.: C, 61.91; H, 5.65; N, 9.63

Found: C, 61.91; H, 5.66; N, 9.61.

EXAMPLE 461,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(carboxymethoxy)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-vi

Compound 17-vi is prepared by hydrolyzing the ester 17-v obtained inExample 45 with an alkali in a conventional manner.

M.p.=257°-259° C.

IR Vx (KBr): 3400, 1755, 1698, 1650, 1620, 1565, 1425 cm⁻¹.

NMR (DMSO- d₆)δ: 0.77-1.03 (8H, m), 2.06-2.22 (2H₇ m), 4.57 (2H, s) 4.86(2H, s), 4.82 (2H, d, J=18.2 Hz), 4.99 (1H, d, J=8.2 Hz), 4.99 (2H, d,J=18.2 Hz), 6.45 (1H, dd, J=6.8 & 2.0 Hz), 6.77-6.90 (2H, m) 7.06 (1H,brs.), 7.11 (1H, t, J=8.0 Hz), 7.28-7,49 (4H, m), 9.15 (1H, s).

Elemental Analysis (for C₂₈ H₂₈ N₄ O₈ ·0.7H₂ O)

Calcd.: C, 59.93; H, 5.28; N, 9.98

Found: C, 59.93; H, 5.12; N, 9.97.

EXAMPLE 471,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(tetrazoylmethoxy)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-vii

1) 3-Nitorphenoxyacetonitlile

To a solution of 3-nitrophenol (6.955 g, 50 mmol) in dimethylformamideare added 97% bromoacetonitrile (6.944 g, 55 mmol), potassium iodide(457 mg, 2.75 mmol) and potassium carbonate (7.60 g, 55 mmol) and themixture is stirred for 15 hr at room temperature. The reaction mixtureis concentrated under reduced pressure and the concentrate ispartitioned between a mixture of ethyl acetate and water. The organiclayer is dried over magnesium sulfate and concentrated under reducedpressure. The resultant crude product is purified by chromatography onsilica gel (toluene/ethyl acetate (9:1)) to give the titled compound(7.128 g; yield, 80%) as white crystals.

2) 3-(Tetrazo-5-yl)methoxyaniline

3-Nitorphenoxyacetonitlile obtained in 1) above is treated with sodiumazide to convert the nitrile group into tetrazole group, followed bycatalytic reduction, in accordance with the method described in EP 0 508796 A1, page 19.

NMR(CD₃ OD)δ: 5.36 (2H, s), 6.39-6.48 (3H, m), 7.05 (1H, t, J=0.8 Hz).

The titled compound is prepared in a manner similar to that described inExample 41 for the preparation of Compound 17i using3-(tetrazo-5-yl)methoxyaniline prepared above and amine 16 h prepared inPreparation 15.

M.p.=203°-204° C.

IR ν_(max) (KBr): 3380, 1700, 1605, 1558, 1502, 1430 cm⁻¹.

NMR (DMSO-d₆)δ: 0.78-1.06 (8H, m), 2.06-2.23 (2H, m), 4.80 (2H, d,J=18.2 Hz), 4.99 (1H, d, J=8.0Hz), 5.00 (2H, d, J=18.2 Hz), 5.40 (2H, s)6.58 (1H, m), 6.81-6.94 (2H, m), 7.10-7.22 (2H, m), 7.26-7,49 (4H, m),9.18 (1H, s).

Elemental Analysis (for C₂₈ H₂₈ N₈ O₆ ·0.5H₂ O)

Calcd.: C, 57.83; H, 5.03; N, 19.27

Found: C, 57.90; H, 4.98; N, 19.24.

EXAMPLE 481,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(carboethoxymethylthio)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-viii

To a solution of the amine above (95 mg) in tetrahydrofuran (5 ml) isadded triphosgene (50 mg) at 0° C. After adding triethylamine (40 μl) in5 portions over 15 min, the mixture is stirred for 5 min at roomtemperature. The mixture is cooled again to 0° C. and a solution ofamine 16 h (145 mg) in tetrahydrofuran (5 ml) is added thereto. Afterstirring for 1 hr at room temperature, ethyl acetate is added to themixture. The reaction mixture is washed with 5% HCl and water in series,dried over sodium sulfate and distilled to remove the solvent.Crystallization of the resultant residue from methylene chloride/ethergives the titled Compound 17-viii (200 mg, 82.6%).

M.p.=193°-196° C.

IR ν_(max) (KBr): 3379, 1716, 1699, 1649, 1609, 1587, 1545 cm⁻¹.

NMR (CDCl₃)δ: 0.87-1.16 (8H, m), 1.20 (3H, t, J=7.2 Hz), 2.00 (2H, m),3.62 (2H, s), 4.14 (2H, q, J=7.2 Hz), 4.77 (2H, d, J=18.0Hz), 4.93 (2H,d, J=18.0 Hz), 5.34 (1H, d, J=7.4 Hz), 6.50 (1H, d, J=7.8 Hz), 6.98 (1H,dt, J=2.2, 6.6 Hz), 7.05-7.18 (2H, m), 7.20-7.36 (5H, m), 7.39 (1H,m).

Elemental Analysis (for C₃₀ H₃₂ N₄ O₇ S·0.2H₂ O)

Calcd.: C, 60.43; H, 5.48; N, 9.40; S, 5.38

Found: C, 60.50; H, 5.55; N, 9.53; S, 5.34.

EXAMPLE 491,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(carboxymethylthio)phenyl)ureido)-1H-l,5-benzodiazepine-2,4(3H,5H)-dione17-ix

Compound 17-ix is prepared by hydrolyzing Compound 17-viii in aconventional manner.

IR ν_(max) (KBr): 3382, 1705, 1673, 1638, 1601, 1589, 1547 cm⁻¹.

NMR (CDCl₃ +CD₃ OD)δ: 0.87-1.20 (8H, m),2.02 (2H, m), 7.00 (1H, dt,J=1.8, 7.4 Hz), 7.14 (1H, t, J=7.6 Hz), 7.23-7.39 (4H, m), 7.42 (1H, t,J=2.0 Hz).

Elemental Analysis (for C₂₈ H₂₈ N₄ O₇ S·0.5H₂ O)

Calcd.: C, 58.68; H, 5.10; N, 9.77 ; S, 5.59

Found: C, 58.49; H, 5.26; N, 10.06; S, 5.63.

EXAMPLE 501,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(5-keto-1,2,4-oxadiazol-3-yl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-x

Compound 17-x is prepared usingm-(5-keto-1,2,4-oxadiazolyl)phenylisocyanate (lit., EP 0508796 A1) andamine 16 h in 53.4% yield.

IR ν_(max) (KBr): 3426, 1789, 1758, 1700,1640 cm⁻¹.

NMR (CDCl₃ +CD₃ OD)δ: 0.90-1.13 (6H, m), 1.19-1.33 (2H, m), 2.03 (2H,m), 4.61 (2H, d, J=18.0Hz), 5.17 (2H, d, J=18.0Hz), 5.45 (1H, d, J=7.8Hz), 6.57 (1H, s), 6.94 (1H, dd, J=2.8, 8.0 Hz), 6.99 (1H, d, J=8.0Hz),7.20 (2H, m),7.33 (1H, m), 7.40 (2H, m), 7.50 (1H, d, J=8.0Hz), 8.28(1H, s), 10.98 (1H, s).

Elemental Analysis (for C₂₈ H₂₆ N₆ O₇ S·0.5H₂ O)

Calcd.: C, 59.26; H, 4.79; N, 14.81

Found: C, 59.25; H, 4.99; N, 14.92.

EXAMPLE 511,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(acetylsulfamoyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-xi

Compound 17-xi is prepared in a manner similar to that described aboveusing m-(acetylsulfamoyl)phenyl-isocyanate (lit., EP 0508796 A1) andamine 16 h in 31.0% yield.

IR ν_(max) (KBr): 3392, 1699, 1664, 1646, 1597, 1553 cm⁻¹.

NMR (DMSO-d₆)δ: 0.80-1.04 (8H, m), 1.86 (3H, s), 2.15 (2H, m), 4.85 (2H,d, J=18.0Hz), 4.99 (2H, d, J=7.8 Hz), 4.99 (1H, d, J=18.0 Hz), 6.91 (1H,d, J=7.8 Hz), 7.27-7.53 (8H, m), 7.99 (1H, s), 9.49 (1H, s).

Elemental Analysis (for C₂₈ H₂₉ N₅ O₈ S·1.7H₂ O)

Calcd.: C, 53.70; H, 5.21; N, 11.18; S, 5.12

Found: C, 53.59; H, 5.08; N, 11.37; S, 5.30.

EXAMPLE 521,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(carbomethoxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-xii

1)3-(N'-(m-(carbomethoxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione

The titled compound is prepared in a manner similar to that describedabove using m-carbomethoxymethyl)phenylisocyanate and amine 4 in 86.8%yield.

M.p.>300° C.

IR ν_(max) (KBr): 3372, 1717, 1678, 1607, 1600, 1559, 1501 cm⁻¹.

NMR (DMSO-d₆)δ: 3.60 (5H, s), 4.63 (1H, d, J=7.4 Hz), 6.80 (1H, d, J=6.6Hz), 6.84 (1H, d, J=7.4 Hz), 7.11-7.31 (7H, m), 9.14 (1H, s), 10.77 (2H,s).

Elemental Analysis (for C₁₉ H₁₈ N₄ O₅ ·0.3H₂ O)

Calcd.: C, 58.58; H, 4.83; N, 14.45

Found: C, 58.70; H, 4.92; N, 14.56.

2) The titled Compound 17-xii is prepared by the N-alkylation carriedout in a manner similar to that described in Examples above. Yield,92.3%; m.p.=199°-202° C.

IR ν_(max) (KBr): 3382, 1699, 1613, 1597, 1558 cm⁻¹.

NMR (CDCl₃) δ: 0.97 (4H, m), 1.10 (4H, m), 2.00 (2H, m), 3.54 (2H, s),3.66 (3H, s), 4.79 (2H, d, J=18.0Hz), 4.91 (2H, d, J=18.0 Hz), 5.33 (1H,d, J=7.4 Hz), 6.42 (1H, d, J=7.8 Hz), 6.89 (1H, dt, J=2.0, 6.2 Hz),7.10-7.36 (8H, m).

Elemental Analysis (for C₂₉ H₃₀ N₄ O₇)

Calcd.: C, 63.73; H, 5.53; N, 10.25

Found: C, 63.54; H, 5.62; N, 10.07.

EXAMPLE 531,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(carbomethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-xiii

Yield, 59.3%; m.p.=209°-211° C.

IR ν_(max) (KBr): 3385, 1700, 1665, 1647, 1616, 1597, 1588, 1501 cm⁻¹.

NMR (DMSO-d₆)δ: 0.80-1.08 (8H, m), 2.15 (2H, qui, J=6.1 Hz), 3.47 (2H,s), 4.84 (2H, d, J=18.0Hz), 4.98 (2H, d, J=18.0Hz), 4.99 (1H, d,J=8.0Hz), 6.80 (1H, dt, J=2.0, 6.8 Hz), 6.85 (1H, d, J=8.0 Hz),7.09-7.47 (7H,m), 9.12 (1H, s), 12.30 (1H, br.s).

Elemental Analysis (for C₂₈ H₂₈ N₄ O₇)

Calcd.: C, 62.31; H, 5.38; N, 10.38

Found: C, 62.59; H, 5.47; N, 10.10.

EXAMPLE 541,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(p-(carbomethoxy)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-xiv

Compound 17-xiv is prepared using, as a starting material,4-methoxyphenylisocyanate obtained from the previously prepared amine 16h and methyl 4-aminobenzoate.

M.p.=278°-279° C.

IR ν_(max) (KBr): 3350, 1721, 1680, 1650, 1600, 1539, 1500, 1435 cm⁻¹.

NMR (DMSO-d₆)δ: 0.78-1.04 (8H, m), 2.06-2.22 (2H, m), 3.79 (3H, s), 4.83(2H, d, J=18.4 Hz), 4.99 (2H, d, J=8.0Hz), 5.00 (2H, d, J=18.4 Hz), 7.03(1H, d, J=7.6 Hz), 7.27-7.53 (4H, m), 7.45 (2H, d, J=8.6 Hz), 7.83 (2H,d, J=8.6 Hz), 9.54 (1H, s).

Elemental Analysis (for C₂₈ H₂₈ N₄ O₇ ·0.4H₂ O)

Calcd.: C, 62.31; H, 5.38; N, 10.38

Found: C, 62.23; H, 5.27; N, 10.40.

EXAMPLE 551,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(p-(carboxy)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-xv

Compound 17-xv is prepared by hydrolyzing the ester 17xiv with an alkaliin a conventional manner.

M.p.=261°-262° C.

IR ν_(max) (KBr): 3400, 1695, 1600, 1545, 1503, 1432 cm⁻¹.

NMR (DMSO-d₆)δ: 0.78-1.05 (8H, m), 2.04-2.23 (2H, m), 4.83 (2H, d,J=18.4 Hz), 5.00 (1H, d, J=7.8 Hz), 5.00 (2H, d, J=18.4 Hz), 7.00 (1H,d, J=7.8 Hz), 7.26-7.51(4H, m), 7.43 (2H, d, J=8.8 Hz), 7.81 (2H, d,J=8.8 Hz), 9.49 (1H, s).

Elemental Analysis (for C₂₇ H₂₆ N₄ O₇ ·0.3H₂ O)

Calcd.: C, 61.19; H, 5.12; N, 10.69

Found: C, 61.85; H, 5.07; N, 10.73.

EXAMPLE 561,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(p-cyanophenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-xvi

Compound 17-xvi is prepared using, as a starting material,4-cyanophenylisocyanate obtained from the previously prepared amine 16 hand 4-cyanoaniline.

M.p.=260°-262° C.

IR ν_(max) (KBr): 3380, 2220, 1703, 1670, 1600, 1540, 1500, 1430 cm⁻¹.

NMR (DMSO-d₆)δ: 0.76-1.05 (8H, m), 2.06-2.24 (2H, m), 4.83 (2H, d,J=18.2 Hz), 4.99 (1H, d, J=7.6 Hz), 5.00 (2H, d, J=18.2 Hz), 7.05 (1H,d, J=8.0 Hz), 7.24-7.63 (4H.sub., m), 7.50 (2H, d,J=8.8 Hz), 7.68 (2H,d, J=8.8 Hz), 9.64 (1H, s).

Elemental Analysis (for C₂₇ H₂₅ N₅ O₅ ·0.4H₂ O)

Calcd.:C, 64.00; H, 5.13; N, 13.82

Found: C, 64.07; H, 5.17;N, 13.65.

EXAMPLE 571,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(m-(tetrazol-4-ylmethylthio)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione17-xvii

1) 3- (Tetrazo-5-yl)methylaniline trifluoroacetate

The titled compound is prepared by treating m-aminobenzenethiol in amanner similar to that described above in connection with thepreparation of 3-(tetrazo-5-yl)methoxyaniline.

NMR(DMSO-d₆)δ: 4.53 (2H, s), 6.72-7.02 (3H, m), 7.10-7.28 (1H, m).

2) Compound 17-xvii is prepared in a manner similar to that described inExample 41 for the preparation of Compound 17i using3-(tetrazo-5-yl)methylaniline trifluoroacetate prepared above 1) and thepreviously prepared amine 16 h.

M.p.=214°-220° C.

NMR (DMSO-d₆)δ: 0.79-1.02 (8H, m), 2.03-2.23 (2H, m), 4.33 (2H, s), 4.83(2H, d, J=18.0 Hz), 4.99 (1H, d, J=8.0 Hz), 5.00 (2H, d, J=18.0 Hz),6.87-7.00 (2H, m), 7.00-7.18 (2H, m), 7.27-7.46 (5H, m), 9.18 (1H, s).

EXAMPLE 581,5-Bis-(pyrrolidinocarbonylmethyl)-3-(N'-(m-(carboethoxy)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione18-i

Compound 18-i is prepared using, as a starting material,3-ethoxycarbonylphenylisocyanate obtained from the previously preparedamine 16a and ethyl 3-aminobenzoate.

NMR (DMSO-d₆)δ: 1.30 (3H, t, J=6.8 Hz), 1.69 2.00 (8H, m), 3.23-3.52(8H, m), 4.29 (2H, q, J=6.8 Hz), 4.49 (2H, d, J=17.2 Hz), 4.72 (2H, d,J=17.2 Hz), 5.00 (1H, d, J=8.0Hz), 6.87 (1H, d, J=8.0Hz), 7.31-7.60 (7H,m), 8.07 (1H, s), 9.49 (1H, s).

Elemental Analysis (for C₄₃ H₅₃ N₇ O₆ ·0.5H₂ O)

Calcd.: C, 65.46; H, 6.96; N, 12.43

Found: C, 65.40; H, 6.91; N, 12.44.

EXAMPLE 591,5-Bis-(pyrrolidinocarbonylmethyl)-3-(N'-(m-(tetrazolylphenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione18-iv

Compound 18-iv is prepared using, as a starting material,3-(1H-tetrazol-5-yl)phenylisocyanate obtained from the previouslyprepared amine 16a and 3-amino-(1H-tetrazol-5-yl)benzene (a compounddisclosed in EP 0 508 796 A1).

M.p.=290°-293° C.

IR ν_(max) (KBr): 3425, 1705, 1642, 1570, 1507, 1455 cm⁻¹.

NMR (DMSO-d₆)δ: 1.64-1,98 (8H, m), 3.23-3.38 (4H, m), 3.42 (4H, m) 4.51(2H, d, J=17.4 Hz), 4.72 (2H, d, J=17.4 Hz), 5.02 (1H, d, J=8.2 Hz),6.81 (1H, d, J=8.2 Hz), 7.19 (1H, d, J=8.2 Hz), 7.35-7.61 (7H, m), 7.82(1H, s), 9.18 (1H, s).

Elemental Analysis (for C₂₁ H₂₇ N₅ O₄ ·0.7H₂ O)

Calcd.: C, 59.20; H, 6.72; N, 16.44

Found: C, 59.38; H, 6.46; N, 16.20.

EXAMPLE 601,5-Bis-(pyrrolidinocarbonylmethyl)-3-(N'-(m-(carbomethoxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione18-xii

The titled Compound 18-xii is prepared in a manner similar to that usedfor the preparation of Compound 17-xii.

IR ν_(max) (KBr): 3425, 1735, 1691, 1652 cm_₁.

NMR (CDCl₃) δ: 1.84 (4H, qui, J=6.2 Hz), 1.97 (4H, qui, J=6.2 Hz),3.36-3.58 (10H, m), 3.67 (3H, s) 4.61 (4H, s), 5.24 (1H, d, J=7.0 Hz),6.44 (1H, d, J=7.4 Hz), 6.87 (1H, dt, J=1.3, 7.4 Hz), 7.13(1H, m),7.20-7.30 (2H, m), 7.30 (2H, m), 7.39 (1H, s), 7.52 (2H, m).

Elemental Analysis (for C₃₁ H₃₆ N₆ O₇ ·9H₂ O)

Calcd.: C, 59.97; H, 6.14; N, 13.54

Found: C, 59.96; H, 6.17; N, 13.54.

EXAMPLE 611,5-Bis-(pyrrolidinocarbonylmethyl)-3-(N'-(m-(carboxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione18-xiii

The titled Compound 18-xiii is prepared by hydrolyzing the compoundprepared in Example 60 in a conventional manner.

M.p.=228°-230° C.

IR ν_(max) (KBr): 3435, 3389, 1705, 1639, 1566 cm⁻¹.

NMR(CD₃ OD)δ: 1.81-2.09 (8H, m), 3.44 (4H, t, J=6.7 Hz), 3.52 (2H, s)3.56 (4H, m), 4.66 (2H, d, J=16.8 Hz), 4.76 (2H, d, J=16.8 Hz), 5.19(1H, s), 6.89 (1H, d, J=7.4 Hz), 7.17 (1H, d, J=7.4, 9.0 Hz), 7.23-7.31(2H, m), 7.41 (2H, m), 7.58 (2H, m).

Elemental Analysis (for C₃₀ H₃₄ N₆ O₇ ·0.7H₂ O)

Calcd.: C, 59.73; H, 5.91; N, 13.93

Found: C, 59.60; H, 5.82; N, 13.97.

Example 621,5-Bis-(cyclopropylmethyl)-3-(N'-(m-tetrazolylphenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione19-iv

Compound 19-iv is prepared using, as a starting material,3-(1H-tetrazol-5-yl)phenylisocyanate obtained from the previouslyprepared amine 16o and 3-amino-(1H-tetrazol-5-yl)benzene (a compounddisclosed in EP 0 508 796 A1).

M.p.=228°-230° C.

IR ν_(max) (KBr): 3380, 1688, 1654, 1600, 1575,1543, 1500, 1425 cm⁻¹.

NMR (DMSO-d₆)δ: 0.03-0.22 (4H, m), 0.22-0.39 (4H,m), 0.71-0.92 (2H, m),3.67 (2H, dd, J=15.8 & 7.0Hz), 4.17 (2H, dd, J=15.8 & 7.0 Hz), 4.83 (1H,d, J=7.8 Hz), 6.94 (1H, d, J=7.8 Hz), 7.38-7.50 (4H, m), 7.52-7.59 (1H,m), 7.66-7.78 (2H, m)), 8.15 (1H, s), 9.43 (1H, s).

Elemental Analysis (for C₂₅ H₂₆ N₈ O₃ ·1.7H₂ O)

Calcd.: C, 58.06; H, 5.73; N, 21.67

Found: C, 58.30; H, 5.44; N, 21.35

EXAMPLE 631,5-Bis-(thienylcarbonylmethyl)-3-(N'-(m-tetrazolylphenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione20-iv

Compound 20-iv is prepared using, as a starting material,3-(1H-tetrazol-5-yl)phenylisocyanate obtained from the previouslyprepared amine 16g and 3-amino-(1H-tetrazol-5-yl)benzene (a compounddisclosed in EP 0 508 796 A1).

M.p.=243°-244° C.

IR ν_(max) (KBr): 3390, 1705, 1663, 1573, 1502,1408, 1240 cm⁻¹.

NMR (DMSO-d₆)δ: 5.15 (1H, d, J=8.0Hz), 5.30 (2H, d, J=17.8 Hz), 5.55(2H, d, J=17.8 Hz), 6.93 (1H, d, J=8.0Hz), 7.27-7.60 (9H, m), 8.01 (1H,s), 8.06-8.20 (4H, m), 9.29 (1H, s).

Elemental Analysis (for C₄₃ H₅₃ N₇ O₆ ·0.5H₂ O)

Calcd.: C, 65.46; H, 6.96; N, 12.43

Found: C, 65.40; H, 6.91; N, 12.44.

EXAMPLE 64 ##STR13##

To a solution of Compound 17-ix (318 mg, 0.56 mmol) prepared in Example49 in dichloromethane (12 ml) and methanol (3 ml) is added 80%m-chloroperbenzoic acid (122 mg, 0.56 mmol) at -5° C. and the mixture isstirred for another 1 hr. To the reaction mixture is added 5% aqueoussodium thiosulfate solution and the mixture is extracted withdichloromethane. The organic layer is washed with water, dried oversodium sulfate and concentrated under reduced pressure. The resultantcrystalline residue is crystallized from dichloromethane-ether to obtainthe titled Compound 17-xix (140 mg; yield, 43%) as white crystals.

M.p.=216°-217° C.

NMR (CDCl₃ +CD₃ ODδ: 0.96-1.18(8H,m), 1.94-2.10 (2H,m), 3.65 (1H, d,J=14 Hz) 3.83 (1H, d, J=14 Hz), 3.83 (1H, d, J=14 Hz), 4.79 (2H, d.d,J=18 & 2.6 Hz), 4.95 (2H, d, J=18 Hz) 7.22- 7.36 (7H, m), 7.67. (1H,S).

EXAMPLE 65 ##STR14##

The S-oxide (250 mg, 0.43 mmol) obtained according to the method aboveis treated with a solution of excessive diazomethane in ether. Theresultant crude product, when purified by chromatography on silica gel(ethyl acetate/methanol (100:0.5)) and crystallized fromdichloromethane-ether, gives the titled Compound 17-xix (200 mg; yield,78%) as white crystals.

M.p.=208°-210° C.

NMR(CDCl₃ +CD₃ OD)δ: 0.95-1.18 (8H, m), 1.88-2.08 (2H,m), 3.68 (1H, d,J=14 Hz), 3.71 (3H, S), 3.82 (1H, d, J=14 Hz), 4.79 (2H, d.d, J=11.4 &6.6 Hz), 4.96 (2H, d, d, J=14.8 & 3.2 Hz), 5.31 (1H, d, J=7.4 Hz), 6.71(1H, d, J=7.4 Hz), 7.13-7.43 (6H,m), 7.52 (1H, d, J=9.4 Hz), 7.62(1H,S), 8.43(1H,S).

A hybrid-type compound was prepared by coupling a compound (I) of thepresent invention prepared in Examples above with an H₂ B. ##STR15##

Reference Example 11,5-Bis-(pyrrolidinecarbonylmethyl)-3-(N'-(3-(3-(3-(3-(pyrrolidinomethyl)phenoxy)propylcarbamoyl)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione12a

A solution of Compound 11a (224 mg, 0.324 mmol) prepared in Example 34,N,N-dimethylaminopropyl-N'-ethylcarbodiimide 2HCl (138 mg, 0.720 mmcl)and 1-hydroxybenzotriazole (54 mg, 0.355 mmol) in dimethylformamide (10ml) is stirred for 30 min under ice-cooling. To the mixture are added asolution of 3-((3-piperidinomethyl)phenoxy)propylamine (130 mg, 0.523mmol) in dimethylformamide (2 ml) and triethylamine (0.2 ml). Themixture is stirred for 16 hr at room temperature and distilled under thereduced pressure to remove the solvent. The resultant residue ispurified by chromatography on silica gel (42 g gel, CHCl₃ /MeOH (9:1))to obtain Compound 12a (198 mg, 66.2%).

IR ν_(max) (KBr): 3416, 1707, 1652, 1559 cm⁻¹.

NMR (CDCl₃) δ: 1.44 (2H, m), 1.62 (4H, m), 1.70-2.04 (12H, m), 2.21 (2H,t, J=6.6 Hz), 2.50 (4H, br.s), 3.17 (2H, m), 3.24-3.50 (1OH, m), 3.55(2H, s), 3.98 (2H, t, J=5.8 Hz), 4.53 (2H, d, J=16.4 Hz), 4.62 (2H, d,J=16.4 Hz), 4.93 (2H, s), 5.19 (1H, d, J=7.0 Hz), 6.07 (1H, m),6.63-6.91 (4H, m), 6.96-7.40 (8H, m), 7.47 (2H, m), 8.44 (1H, s).

Elemental Analysis (for C₄₉ H₆₃ N₉ O₉ ·1.5H₂ O)

Found: C, 62.05; H, 6.93; N, 13.31

Calcd.: C, 62.01; H, 7.01; N, 13.28.

Compounds described in the following Reference Examples are prepared ina manner similar to that described in Reference Example 1.

Reference Example 21,5-Bis-(cyclopropylcarbamoylmethyl)-3-(N'-(3-(3-(3-(3-(pyrrolidinomethyl)phenoxy)propylcarbamoyl)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione12c

IR ν_(max) (KBr): 3360, 3316,1708, 1665, 1599, 1556 cm⁻¹.

NMR (CDCl₃ +CD₃ OD)δ: 0.44 (4H, m), 0.65 (4H, m), 1.48 (2H, m), 1.66(4H,m), 1.80 (2H, m), 1.96 (2H, m), 2.40-2.70 (6H, m), 3.16 (2H, t,J=6.5 Hz),3.38 (2H, m), 3.60 (2H, br.s), 4.02 (2H, t, J=6.2 Hz), 4.33(2H, d, J=16.2 Hz), 4.71 (2H, d, J=16.2 Hz), 5.00 (2H, s), 5.11 (1H, s),6.81 (1H, dd,J=7.9, 1.7 Hz), 6.89(2H, m), 6.99 (1H, s), 7.13-7.31 (4H,m), 7.37 (4H,s).

Elemental Analysis(For C₄₇ H₅₉ N₉ O₉ ·1.2H₂ O)

Found: C, 61.63; H, 6.71; N, 13.76

Calcd.: C, 61.65; H, 6.76; N, 13.77.

Reference Example 31,5-Bis-(o-methylphenacyl)-3-(N'-(3-(3-(3-(3-piperidinomethyl)phenoxy)propylcarbamoyl)-propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione12d

M.p.=132°-133° C.

IR ν_(max) (nujol): 3347, 1702, 1690, 1668, 1612, 1568 cm₁.

NMR (DMSO-d₆) δ: 1.31-1.55 (6H, m), 1.61 (2H, quintet, J=7.0 Hz), 1.81(2H, quintet, J=6.0Hz), 2.06 (2H, t, J=7.8 Hz), 2.26-2.39 (4H, m), 2.36(6H,s), 2.97 (2H, q, J=6.0Hz), 3.18 (2H, q, J=6.0 Hz), 3.34 (2H,s), 3.94(2H, t, J=6.4 Hz), 4.92 (2H, s), 5.07 (1H, d, J=8.0Hz), 5.09 (2H, d,J=17.6 Hz), 5.20 (2H, d, J=17.6 Hz), 6.71-7.00 (5H, m), 7.12-7.58 (15H,m), 7.73-7.95 (3H, m), 9.16 (1H, s).

Elemental Analysis (for C₅₅ H₆₁ N₇ O₉ ·0.5H₂ O)

Found: C, 67.90; H, 6.35; N, 10.05

Calcd.: C, 67.88; H, 6.42; N, 10.08.

Reference Example 41,5-Bis-(2-furylcarbonylmethyl)-3-(N'-(3-(3-(3-piperidinomethyl)phenoxy)propylcarbamoyl)-propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione12f

M.p.=140°-145° C.

IR ν_(max) (nujol): 1705, 1691, 1639, 1598, 1571 cm_₁.

NMR (DMSO-d₆)δ: 1.31-1.55 (6H, m), 1.61 (2H, quintet, J=7.0Hz),1.81 (2H,quintet, J=6.0Hz), 2.06 (2H, t, J=7.8 Hz), 2.26-2.39 (4H, m), 2.97 (2H,q, J=6.0Hz), 3.18 (2H, q, J=6.0 Hz),3.34 (2H, s), 3.94 (2H, t, J=6.0Hz),4.92 (2H, s), 5.11 (1H,d, J=8.0Hz), 5.12 (2H, d, J=8.0Hz), 5.38 (2H, d,J=18.0Hz), 6.80 (2H, d.d, J=3.6 & 1.8 Hz), 6.80-6.93 (5H, m), 7.13- 7.54(9H, m), 7.65 (2H, d.d, J=3.6 & 0.6 Hz), 7.88 (1H, t, J=5.8 Hz), 8.10(2H, d.d, J=1.8 & 0.6 Hz), 9.18 (1H, s).

Elemental Analysis (for C₄₉ H₅₃ N₇ O₁₁ ·H₂ O)

Found: C, 63.06;H, 5.83; N, 10.56

Calcd.: C, 63.01; H, 5.94; N, 10.50.

Reference Example 51,5-Bis-(2-thienylcarbonylmethyl)-3-(N'-(3-(3-(3-(3-(piperidinomethyl)phenoxy)propylcarbamoyl)-propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione12g

M.p.=165°-167° C.

IR ν_(max) (nujol): 3338, 1704, 1666, 1636, 1567 cm⁻¹.

NMR (DMSO-d6) δ: 1.31-1.55 (6H, m), 1.61 (2H, quintet, J=7.0Hz),1.81(2H, quintet, J=6.0Hz), 2.06 (2H, t, J=7.8 Hz), 2.26-2.39 (4H, m), 2.97(2H, q, J=6.0Hz), 3.18 (2H, q, J=6.0Hz),d.34 (2H, s), 3.94 (2H, t,J=6.0Hz), 4.92 (2H, s), 5.12 (1H,d, J=8.0 Hz), 5.29 (2H, d, J=18.1 Hz),5.54 (2H, d, J=18.1 Hz), 6.70-6.97 (5H, m), 7.15-7.39 (7H, m), 7.40-7.50(4H, m), 7.88 (1H, t, J=6.0 Hz), 8.07-8.18 (4H, m), 9.18 (1H, s).

Elemental Analysis (for C₄₉ H₅₃ N₇ O₉ S₂ ·0.8H₂ O)

Found: C, 61.14; H, 5.80; N, 10.38

Calcd.: C, 61.14; H, 5.72; N, 10.19; S, 6.66.

Reference Example 6 1,5-Bis-(cyclopropylcarbonylmethyl)-3-(N'-(3-(3-(3-(3-(piperidinomethyl)phenoxy)propylcarbamoyl)-propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione12 h

Yield, 71.6%.

IR ν_(max) (KBr): 3357, 1701, 1598, 1559, 1502 cm⁻¹.

NMR (CDCl₃)δ: 0.95 (4H, m), 1.06 (4H, in), 1.57 (4H, m), 1.70-2.05 (6H,m), 2.18 (2H, t, J=6.8 Hz), 2.42 (4H, br.s), 3.17 (2H, qui, J=5.4 Hz),3.36 (2H, q, J=6.0 Hz), 3.47 (2H, s), 3.96 (2H, t, J=5.8 Hz), 4.75 (2H,d,J=18.0 Hz), 4.89 (2H, d, J=18.0 Hz), 4.95 (2H, s), 5.31 (1H, d, J=7.6Hz), 5.71 (1H, t, J=5.3 Hz), 6.68-6.79 (6H, m), 7.03-7.35 (8H, m), 8.06(1H, s).

Elemental Analysis (for C₄₇ H₅₇ N₇ O₉ ·0.7H₂ O)

Found: C, 64.41; H, 6.78; N, 11.32

Calcd.: C, 64.40; H, 6.71; N, 11.18.

Reference Example 71,5-Bis-(cyclopentylcarbonylmethyl)-3-(N'-(3-(3-(3-(3-(piperidinomethyl)phenoxy)propylcarbamoyl)-propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione12i

Yield, 68.5%.

IR ν_(max) (KBr): 3398, 1702, 1670, 1646, 1614, 1599, 1501 cm⁻¹.

NMR (CDCl₃)δ: 1.35-2.00 (26H, m), 2.20 (2H, t, J=6.6 Hz), 2.45 (4H,br.s), 2.91 (2H, qui, J=7.9 Hz), 3.18 (2H, q, J=5.7 Hz), 3.36 (2H, q,J=6.0 Hz), 3.50 (2H, s), 3.96 (2H, t, J=5.8 Hz), 4.66 (2H, d, J=18.0Hz),4.76 (2H, d, J=18.0Hz), 4.96 (2H, s), 5.28 (1H, d, J=7.0Hz), 5.71 (1H,t, J=6.0Hz), 6.65 (1H, d, J=7.6 Hz), 6.61-6.90 (4H, m), 6.97 (1H, s),7.06-7.39 (8H, m), 8.06 (1H, s).

Elemental Analysis (for C₅₁ H₆₅ N₇ O₉ ·H₂ O)

Found: C, 65.23; H, 7.14; N, 10.59

Calcd.: C, 65.30; H, 7.20; N, 10.45.

Reference Example 81,5-Bis-(l-methylimidazol-2-ylmethyl)-3-(N'-(3-(3-(3-(3-(piperidinomethyl)phenoxy)-propylcarbamoyl)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione12j

Yield, 77.1%.

NMR (CDCl₃)δ: 1.44 (2H, m), 1.58 (4H, m), 1.78 (2H, m), 1.91 (2H, m),2.17 (2H, t, J=6.6 Hz), 2.42 (4H, br.s), 3.15 (2H, qui, J=5.8 Hz), 3.36(2H, q, J=6.2 Hz), 3.48 (2H, s), 3.50 (6H, s), 3.96 (2H, t, J=5.8 Hz),4.94 (2H, s), 4.98 (4H, s), 5.14 (1H, d, J=7.4 Hz), 5.77 (1H, t, J=6.4Hz), 6.65-6.98 (6H, m), 6.73 (2H, d, J=1.2 Hz), 6.88 (2H, d, J=1.2 Hz),7.04-7.25 (4H, m), 7.30 (2H, m), 7.77 (2H, m), 8.42 (1H, s).

Elemental Analysis (for C₄₇ H₅₇ N₁₁ O₇ ·13H₂ O)

Found: C, 61.93; H, 6.64; N, 17.13

Calcd.: C, 61.94; H, 6.59; N, 16.90.

Reference Example 91,5-Bis-allyl-3-(N'-(3-(3-(3-(3-(piperidinomethyl)phenoxy)propylcarbamoyl)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione12n,

Amorphous solid.

IR ν_(max) (nujol): 3308, 1697, 1665, 1638, 1614, 1565 cm⁻¹.

NMR (DMSO-d₆)δ: 1.31-1.55 (6H, m), 1.61 (2H, quintet, J=7.0Hz), 1.81(2H,quintet, J=6.0 Hz), 2.06 (2H, t, J=7.8 Hz), 2.26-2.39 (4H, m), 2.97(2H, q, J=6.0 Hz), 3.18 (2H, q, J=6.0Hz), 3.34 (2H,s), 3.94 (2H, t,J=6.4 Hz), 4.45 (2H, d.d, J=16.0 & 5.8 Hz), 4.67 (2H, d.d, J=16.0 & 5.8Hz), 4.90 (1H, d, J=7.4 Hz), 4.92 (2H, s),5.08 (2H, s), 5.14 (2H, d,J=3.6 Hz), 5.59-5.81 (2H, m), 6.72-6.95 (5H, m), 7.12-7.30 (4H, m), 7.35(1H, s), 7.36-7.48 (2H, m), 7.55-7.68 (2H, m), 7.87 (1H, t, J=5.4 Hz),9.21 (1H, s).

Elemental Analysis (for C₄₃ H₅₃ N₇ O₆ ·0.5H₂ O)

Found: C, 65.40; H, 6.91; N, 12.44

Calcd.: C, 65.46; H, 6.96; N, 12.43.

Reference Example 101,5-Bis-(cyclopropylmethyl)-3-(N,-(3-(3-(3-(3-(piperidinomethyl)phenoxy)propylcarbamoyl)-propylcarbamoyloxymethyl)phenyl)ureido)-1H-l,5-benzodiazepine-2,4(3H,5H)-dione12o

Amorphous solid.

IR ν_(max) (nujol): 3383, 3297, 1686, 1658, 1637, 1604, 1569 cm⁻¹.

NMR (DMSO-d₆)δ: 0.09-0.18 (4H, m), 0.23-0.37 (4H, m), 0.72-0.92 (2H, m),1.31-1.55 (6H, m), 1.61 (2H, qui, J=7.0Hz), 1.81 (2H, quintet, J=6.0Hz),2.06 (2H, t, J=7.8 Hz), 2.24-2.33 (4H, m), 2.97 (2H, q, J=6.0Hz), 3.18(2H, q, J=6.0Hz), 3.34 (2H,s), 3.66 (2H, d.d, J=14.0 & 7.0 Hz), 3.94(2H, t, J=6.0 Hz), 4.16 (2H, d.d, J=14.0 & 7.0 Hz), 4.79 (1H, d, J=7.4Hz), 4.91 (2H, s), 6.72-6.91 (5H, m), 7.12-7.29 (4H, m), 7.36 (1H, s),7.38-7.49 (2H, m), 7.67-7.78 (2H, m), 7.86 (1H, t, J=5.6 Hz), 9.21 (1H,s).

Elemental Analysis (for C₄₅ H₅₇ N₇ O₇)

Found: C, 66.87; H, 7.14; N, 12.08

Calcd.: C, 66.89; H, 7.11; N, 12.13.

Reference Example 111,5-Bis-(tert-butyloxycarbonyl-methyl)-3-(N'-(3-(3-(3-(3-(piperidinomethyl)phenoxy)-propylcarbamoyl)propylcarbamoyloxymethyl)phenyl)ureido)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione12s

Yield, 64.1%.

IR ν_(max) (KBr): 3381, 1741, 1711, 1677, 1646, 1614, 1599, 1560 cm⁻¹.

NMR (CDCl₃)δ: 1.43 (20H, m), 1.59 (4H, m), 1.73-2.00 (4H, m), 2.19 (2H,t, J=7.0Hz), 2.45 (4H, br.s), 3.20 (2H, qui, J=5.8 Hz), 3.35 (2H, q,J=6.1 Hz), 3.50 (2H, s), 3.97 (2H, t, J=6.0Hz), 4.38 (2H, d J=17.6 Hz),4.52 (2H, d, J=17.6 Hz), 4.95 (2H, s), 5.08 (1H, br.s), 5.31 (1H, d,J=7.6 Hz), 5.60 (1H, t, J=6.0Hz), 6.70-7.23 (9H, m), 7.36 (4H, s), 7.89(1H, s).

Elemental Analysis (for C₄₉ H₆₅ N₇ O₁₁ ·0.8H₂ O)

Found: C, 62.38; H, 7.08; N, 10.50

Calcd.: C, 62.44; H, 7.12; N, 10.40.

Compounds of the formula (I) of the present invention prepared inExamples were subjected to in vitro and in vivo tests for evaluating theantagonistic activity gastrin receptor and/or CCK-B receptor.

Experiment 1 In Vivo Test Evaluation of Inhibitory Effect on GastricAcid Secretion by Schild Method

Twenty four hour starved (ad libitum for water) male Sprague Dawley rats(8-week-old) were anesthetized with urethane (1.5 g/kg S.C.) and keptbreathing with esophagus cannulas. After laparotomy, esophagus cannulaswere inserted orally up to proventriculus and ligated around gastriccardiac. Perfusion cannulas were inserted from duodenum into stomach andligated around pylorus. Another cannulas were placed into duodenum andligated for administration of drug. After sutra of abdomen, stomach wasperfused via the esophagus cannulas with physiological saline (37° C.)while collecting perfusate for a 15 min interval. The perfusate wassubjected to titration with 0.001 N NaOH solution to determine theacidity. When the basal acid secretion became stable, pentagastrin (10μl/kg/hr) was administered in a sustained manner via common carotid veinfor about 90 min until the acid secretion reached approximately thehighest level, when a test compound (0.5% M.C. suspension) wasadministered into duodenum through cannulas. The perfusate was collectedfor a 15 min interval to monitor the acid secretion for 90 min. Thepercent inhibition was calculated as follows:

    Percent inhibition (%)=100×(A-B)/(C-B)

A: the minimum value of total acidity observed after the administrationof a test compound;

B: the total acidity obtained immediately before the administration ofpentagastrin; and

C: the total acidity obtained immediately before the administration of atest compound.

Results are summarized in Table 1 below.

Experiment 2 In Vitro Test for Evaluation of Gastrin and/or CCK-BAntagonism

The pharmacological effect of compounds (I) prepared in Examples abovewere evaluated in vitro with respect to antagonistic activity againstgastrin receptor, CCK-B receptor or CCK-A receptor, using fundic glandcells of guinea pig, crude membrane specimen from mouse cerebral cortex,or crude membrane specimen from mouse pancreas, respectively.

Animals used in test

Male Hartley guinea pig (450-600 g) or male ddY mouse (24-30 g) wereused.

(1) Gastrin Receptor Antagonism

Preparation of gastric glands:

Male Hartley guinea pigs (450-600 g) were killed by bleeding and stomachwas extracted from each animal immediately, from which gastric glandswere prepared.

Preparation of test compounds and procedures of displacing assay

A 1M solution of a compound to be tested in DMSO is prepared and dilutedwith 50% DMSO to obtain a ten-fold dilution series.

The reaction is initiated by the addition of gastric glands to solutionsof different concentration each containing ¹²⁵ I-labeled gastrin (finalconcentration, 0.2 nM). The mixture is incubated for 30 min at 25° C.,centrifuged at 2000 rpm for 5 min and the supernatant is removed byaspiration. To the pellet is added ice-cooled incubation buffer andmixed gently, followed by an immediate centrifugation and removal of thesupernatant by aspiration. The radioactivity is counted on gammacounter. The same procedure was repeated using 50% DMSO solution orhuman gastrin I (final concentration, 2 μM) instead of a solution oftest compound so as to obtain the control value regarding total bindingor the value regarding non-specific binding, respectively.

Calculation of IC₅₀ :

The IC₅₀ was determined by plotting the ratio (%) of specific binding ofa test compound to that of control on semilogarithmic graph andobtaining the concentration corresponding to 50%, wherein:

specific binding of control=total binding (cpm)-non-specific binding(cpm); and

specific binding of test compound=total binding (cpm)-non-specificbinding (cpm).

(2) CCK-A Receptor Antagonism and CCK-B Receptor Antagonism

Preparation of CCK receptor preparations

Male ddY mice (24 to 30 g) were killed by decapitation and cerebralcortex (CCK-B) and pancreas (CCK-A) were extracted immediately. Each ofcerebral cortex and pancreas was mixed with 50 mM Tris-HCl buffer (pH7.4) and homogenized with a teflon-glass homogenizer and polytronhomogenizer to obtain crude membrane specimens.

Preparation of test compounds and procedures of displacing assay A 1Msolution of a compound to be tested in DMSO is prepared and diluted with50% DMSO to obtain a ten-fold dilution series.

The reaction is initiated by the addition of crude membrane specimen tosolutions of different concentration each containing ³ H!CCk-8 (finalconcentration, 1 nM). The mixture is incubated for 90 min at 25° C.,filtered through glass filter with aspiration and washed with a cooled50 mM Tris buffer. After the addition of Aquazol-2 cocktail theradioactivity is counted. The same procedure was repeated using 50% DMSOsolution or Ceruletide (final concentration, 1 μM) instead of a solutionof test compound so as to obtain the control value regarding totalbinding or the value regarding non-specific binding, respectively.

Calculation of IC₅₀ :

The IC₅₀ was determined by plotting the ratio (%) of specific binding ofa test compound to that of control on semilogarithmic graph andobtaining the concentration corresponding to 50%, wherein:

specific binding of control=total binding (cpm)-non-specific binding(cpm); and

specific binding of test compound=total binding (cpm)-non-specificbinding (cpm).

Results are shown in Table 1 below.

                  TABLE 1                                                         ______________________________________                                                                 Inhibitory Effect                                            Receptor         on acid secretion                                            Gastrin                                                                              CCK-B    CCK-A    (Schild method)                              Compound No.                                                                            (IC.sub.50, nM)    (mg/kg):(%)*                                     ______________________________________                                        8a       7         54       860    0.3:49                                     8d       4         11       29     0.3:66                                     8f       3         17       120    0.3:73                                     8h       5         18       380    0.3:81                                     8o       12        23       860    0.3:60                                     8r       18        21       760    0.3:46                                     8s       2         2        100                                               9a       8         130      2,100                                             9b       14        120      360                                               9d       6         11       <100                                              9f       7         62       1,400                                             9g       3         7        420                                               9h       4         20       1,250                                             9i       7         2        290                                               9o       31        78       3,900                                             10c      64        380      2,100                                             10s      7         4        <100                                              11f      6         38       800                                               11o      18        36       1,850                                             17-i     7         140      1,350  0.3:44                                     17-ii    5         105      >10,000                                                                              0.3:72                                     17-iii   6         190      11,500 0.3:47                                     17-iv    2         8        2,900  0.1:44                                     17-v     16        180      3,600  0.3:59                                     17-vi    3         16       4,400  0.3:22                                     17-vii   11        4        1,800  0.3:68                                     17-viii  2         6        460    0.3:76                                     17-ix    0.6       2        500    0.1:74                                     17-x     1         3        360    0.3:70                                     17-xi    6         115      >10,000                                                                              0.3:39                                     17-xii   15        48       2,500  0.3:48                                     17-xiii  2         7        4,800  0.3:21                                     17-xvii  <1        1        1,300  0.1:30                                     18-i     25        440      3,200                                             18-iv    4         38       3,600  0.3:65                                     18-xiii  7         29       10,500 0.3:26                                     19-iv    6         26       2,600  0.5:58                                     20-iv    2         3        380    0.3:65                                     ______________________________________                                    

Experiment 3 In Vitro Test for Evaluation of Histamine H₂ ReceptorAntaqonism

The Histamine H₂ receptor antagonism was evaluated by determining PA₂ inthe following manner.

Male Hartley guinea pigs (450 to 600 g) were killed by bleeding andright atrium was extracted from each animal and suspended in Magnusdevice (Krebs bicarbonate buffer aerated with 95% O₂ and 5% CO₂ at 30°C.). To the device was added histamine cumulatively and the time-courseeffect of histamine was evaluated.

The pA₂ values which are the parameter reflecting the activity ofhistamine H₂ antagonism were calculated as a negative logarithm of aconcentration of an antagonist required to shift the dose-response curveof histamine so that the concentration is doubled. TAKAYANAGI K.,IYAKUHIN-KAIHATSU KISO-KOZA V, (TSUDA K., NOGAMI, T. Ed.), Chijin-Kan,p. 731-776, Tokyo (1974); and Van Rossum, J. M., Arch. Int. Pharmacodyn.Ther., 143: 299-330 (1963)!

Experiment 4 Effect of Gastrin Receptor in the Prevention of Relapse ofUlcer Followinq the Administration of H₂ B

The usefulness of benzodiazepine derivatives prepared in Examples in theanti-ulcer treatment was evaluated in the following experiments. Thus,the effect of combined formulation of a histamine H₂ receptor antagonist(H₂ B) and a known gastrin receptor antagonist in the relief orprevention of the relapse of ulcer following the continuousadministration of H₂ B was evaluated. In the experiment, the evaluationof the inhibitory effect of gastrin receptor antagonist was conductedafter an interruption of few days following a continuous administrationof H₂ B which is thought to be the cause of relapse of ulcer using ascriterion, (1) increase in gastric acid secretion (rebound effect); and(2) the decrease in the protecting function of gastric mucosa.

Materials

Male Sprague Dawley rats weighing from 240 to 280 g were used asexperimental animals.

As H₂ B, famotidine, and as gastrin receptor antagonist, L-365,260described in Example 281 of Japanese Patent Publication (KOKAI)63-238069 (EP 167,919; EP 284,256; U.S. Pat. No. 4,820,834; U.S. Pat.No. 5,004,741)), were used. A combined formulation was prepared bymixing famotidine (M.W., 337.4) and L-365,260 (M.W., 398.44) in theratio of 1:1 because these substances have almost the same molecularweight. For administration, a suspension of 10 mg famotidine orL-365,260 in 1 ml vehicle (0.5% methyl cellulose solution) was prepared.The combined formulation is a mixture of an equal amount of eachsuspension.

Administration Procedure

A previously determined dose of each test compound (1.0 ml or 3.0 ml/kg)was charged in a syringe and administered orally to a rat directly intogaster using a stainless probe needle (φ1.2×L 80 mm) equipped to thesyringe.

Experimental Procedure

A. Single administration test

(1) Gastric damage due to stress induced by restricted water-immersion(single administration test)

Twenty four hour starved (ad libitum for water) male Sprague Dawley rats(270-290 g) were orally administered with a test compound and, 30 minlater, were loaded with stress in the following manner. Rats were placedin a stress cage (Natsume Seisakusyo) and immersed in water tank at 23°C. upto the level of pectoral processus xiphoideus Takagi et al, Chem.Pharm. Bull (Tokyo) 12, 465-472 (1964)!. Seven hours later, animals werewithdrawn from the water tank and killed with ether. The gaster wasextracted and 1% formalin solution (13 ml) was injected in it. Thegaster was fixed by dipping in 1% formalin solution for 10 min, cut outalong the greater curvature and developed on a glass plate (hereinafter,referred to as formalin treatment). The gaster was observed underanatomic microscope (x10) and longer diameter (mm) of each damages(bleeding erosion) appearing around fundic glands was measured.

B. Continuous administration test

(A) A group consists of animals that received vehicle (0.5% methylcellulose solution, 1 ml/kg).

(B) A group consists of animals that received famotidine (a suspensionof 10 mg famotidine in 1 ml 0.5% methyl cellulose) continuously at adose of 10 or 30 mg/kg/day.

(C) A group consists of animals that received L-365,260 continuously ata dose of 10 mg/kg/day.

(D) A group consists of animals that received a combined formulation offamotidine and L-365,260 at a dose of 10 mg/kg/day for each compound.

(2) Basic gastric acid secretion after interruption following thecontinuous administration of famotidine

To a rat bred normally was administered continuously famotidine at adose of 10 or 30 mg/kg/day for a week. After the final administration,the basis gastric acid secretion (total acid excretion) was determinedaccording to the pylorus ligation method (4-hour-method) Shay, H. etal., Gastroenterology, 5: 43-61 (1945)!. The determination was carriedout following the final administration, immediately, 24 and 48 hr (2days) later, in the case of the group regarding continuousadministration of 10 mg/kg/day, and immediately, 3 and 4 days later, inthe case of the group regarding continuous administration of 30mg/kg/day.

Thus, rats were subjected to laparotomy under ether anesthetization andpylorus was ligated. Four hours later, gaster was extracted under etheranaesthetization and the accumulated gastric juice was collected. Aftercentrifugation (3000 rpm, 10 min), pH and acidity of gastric juice wasdetermined. The acidity was measured upto pH 7.0 with 0.1N NaOH. Thetotal acid secretion (μEq/4 hr) was calculated by multiplying acidity bythe volume of juice for each animal.

(3) Aspirin-induced gastric mucosa damage following a continuousadministration of famotidine

After two-day-interruption following a continuous administration offamotidine, aspirin (200 mg/kg) was administered to rats orally. Sevenhours later, gaster was extracted under ether anesthetization andsubjected to formalin treatment with 1% formalin solution. The length(mm) of gastric mucosa damage appearing around fundic glands was thenmeasured. Lesion index was calculated by adding all the measurements.Rats had been starved for 24 hr before aspirin administration (adlibitum for water).

Experimental Results

(1) Gastric damage due to stress induced by restricted water-immersion

Results are shown in Table 2 below.

                  TABLE 2                                                         ______________________________________                                        Effect of famotidine, L-365, 260 and famotidine + L365, 260 on                gastric damage due to stress induced by restricted water-                     immersion in rats                                                                      dose     administration                                                                           rat    lesion index                              Treatment                                                                              (mg/kg)  route      number (% to control)                            ______________________________________                                        Control            p.o.**    10     100 ± 8.1                              (0.5% M.C.)                                                                   Famotidine                                                                             1        p.o.       10     33.4 ± 5.5*                            L-365, 260                                                                             1        p.o.       10     130.4 ± 20.6                           Famotidine +                                                                           1 + 1    p.o.       10      37.8 ± 10.2*                          L-365, 260                                                                    Control           p.o.       10      100 ± 14.7                            (0.5% M.C.)                                                                   Famotidine                                                                             3        p.o.       10     33.4 ± 5.5*                            L-365, 260                                                                             3        p.o.       10     84.7 ± 17.4                            Famotidine +                                                                           3 + 3    p.o.       10     28.2 ± 6.9*                            L-365, 260                                                                    ______________________________________                                         *p < 0.05 (compared with each control)                                        **p.o. = oral administration                                             

L-365,260 does not inhibit the appearance of stress gastric damage ateither dose (1 or 3 mg/kg). Famotidine inhibited the appearancesignificantly at a dosage of 3 mg/kg. The group related to a combinedformulation showed stronger inhibition compared to a single formulationat any dosage. (see, Table 2)

(2) Basic gastric acid secretion after interruption following thecontinuous administration of famotidine

                  TABLE 3                                                         ______________________________________                                        Gastric acid excretion immediately after the continuous                       administration (7 days) of famotidine in rats with pylorus                    ligation                                                                                                  total acid                                                 dose      rat      secretion                                                                              inhibition                               Treatment                                                                              (mg/kg/day)                                                                             number   (μEg/49 hr)                                                                         (%)                                      ______________________________________                                        Control            5        395.4 ± 41.9                                   (0.5% M.C.)                                                                   Famotidine                                                                             10        5        151.8 ± 29.8**                                                                      61.6                                              30        5        158.4 ± 54.9**                                                                      59.9                                     ______________________________________                                         **P < 0.01 (compared with each control)                                  

                  TABLE 4                                                         ______________________________________                                        Gastric acid excretion at 24 and 48 hours after the                           continuous administration (7 days) of famotidine in rats                      with pylorus ligation                                                                                      total acid                                                                              in-                                    Treatment  dose      rat     secretion hibition                               Hr**           (mg/kg/day)                                                                             number                                                                              (μEq/4 hr)                                                                           (%)                                  ______________________________________                                        24   Control             4     593.7 ± 129.9                                    (0.5% M.C.)                                                                   Famotidine                                                                              10        4     821.0 ± 214.2                                                                        -38.3                                48   Control             4     461.2 ± 56.6                                     (0.5% M.C.)                                                                   Famotidine                                                                              10        4      948.9 ± 187.5*                                                                      -105.7                               ______________________________________                                         *P < 0.5 (compared with each control)                                         **After treatment.                                                       

                  TABLE 5                                                         ______________________________________                                        Gastric acid excretion on 3 and 4 days after the continuous                   administration (7 days) of famotidine in rats with pylorus                    ligation                                                                                            rat    total acid                                                                              in-                                    Treatment   dose      num-   secretion hibition                               Day**           (mg/kg/day)                                                                             ber  (μEq/4 hr)                                                                           (%)                                  ______________________________________                                        3     Control             11   585.8 ± 94.2                                      (0.5% M.C.)                                                                   Famotidine                                                                              30         8   850.9.0 ± 176.2                                                                      -45.3                                4     Control             10   473.3 ± 61.2                                      (0.5% M.C.)                                                                   Famotidine                                                                              30        10   682.5.9 ± 65.3*                                                                      -44.2                                ______________________________________                                         *P < 0.5 (compared with each control)                                         **After treatment.                                                       

In the group that received famotidine (10 mg/kg/day) continuously, thebasic gastric acid secretion was inhibited significantly (P<0.05)immediately after interrupting the administration though, it increasedsignificantly (P<0.05) after a 48-hour-interruption. In the group thatreceived famotidine (30 mg/kg/day) continuously, the basic gastric acidsecretion was inhibited significantly (P<0.05) immediately afterinterrupting the administration though, it increased significantly(P<0.05) after a 4-day-interruption. (see, Tables 3, 4 and 5)

                  TABLE 6                                                         ______________________________________                                        Gastric acid excretion on 2 and 3 days after the continuous                   administration (7 days) of famotidine, L-365, 260 and                         famotidine + L-365, 260                                                                                         total acid                                  Treatment     dose       rat      secretion                                   Day**             (mg/kg/day)                                                                              number (μEq/4 hr)                             ______________________________________                                        2      Control               5      609.5 ± 60.5                                  (0.5% M.C.)                                                                   Famotidine 10         5      788.4 ± 217.7                                 L-365, 260 10         4      631.4 ± 147.9                                 Famotidine +                                                                             10 + 10    5      559.0 ± 66.7                                  L-365, 260                                                             3      Control               4      355.8 ± 49.9                                  (0.5% M.C.)                                                                   Famotidine 10         5      682.4 ± 132.6                                 L-365, 260 10         4      681.0 ± 142.9                                 Famotidine +                                                                             10 + 10    5      494.8 ± 130.5                                 L-365, 260                                                             ______________________________________                                         **After treatment.                                                       

With respect to plasma gastrin concentration, it increased significantly(P<0.05) immediately after the continuous administration, reflecting theinhibition of acid excretion compared with control though, only littledifference could be observed between the treated group and control groupon 1 to 3 days after interruption of administration.

The increase in the acid excretion after interruption following thecontinuous administration of famotidine (10 mg/kg/day) had a tendency tobe inhibited by combining famotidine (10 mg/kg) with L-365,260 (10mg/kg).

In the group that received a continuous administration of L-365,260alone for a week, the results did not differed from those obtained incontrol group. (see, Table 6)

(3) Aspirin-induced gastric mucosa damage following a continuousadministration of famotidine

                  TABLE 7                                                         ______________________________________                                        Effect of a continuous administration (7 days) of                             famotidine, L-365, 260 and famotidine + L-365, 260 on aspirin-                induced gastric mucosa damage                                                          dose    administration                                                                            rat   lesion index                               Treatment                                                                              (mg/kg) route       number                                                                              (mm.sup.2)                                 ______________________________________                                        Control          p.o.        5     20.8 ± 4.0                              Famotidine                                                                             10      p.o.        5      67.0 ± 14.2*                           L-365, 260                                                                             10      p.o.        5     43.2 ± 13.7                             Famotidine +                                                                           10 + 10 p.o.        5     27.2 ± 12.2                             L-365, 260                                                                    ______________________________________                                         Note: Aspirin (200 mg/kg) was administered orally to a rat after              twoday-interruption following each continuous administration.                 *P < 0.01 (compared with each control)                                   

Many linear-shaped damages caused by aspirin appeared at the gastricbody.

As is apparent from the table above, in the group that received acontinuous administration of famotidine, the condition deterioratedsignificantly (P<0.05) compared with control group, while in the groupthat received a combined formulation of famotidine and L-365,260, thedeterioration was inhibited significantly (P<0.05). In the group thatreceived L-365,260 continuously, the condition seemed to have tendencyto deteriorate, but it is not significant.

An increase in the basic acid secretion (acid rebound) was observed onthe 2nd day after interruption following the continuous administrationof famotidine. It is considered to be a phenomenon common to H₂ B. Thegastric mucosa reactivity in the group that received continuousadministration of famotidine was tested, which revealed that the ulcerindex got worse significantly on the 2nd day after interruption. Thisresult is in good agreement with the point where the acid rebound wasobserved as mentioned above. These facts indicate that famotidinetreatment weakens the protective function of gastric mucosa.

It was proved that the reverse effect of famotidine, that is, acidrebound phenomenon and deterioration of aspirin-induced gastric mucosadamage, can be inhibited by using famotidine in combination withL-365,260. These results generally show that the appearance of gastricdamages following the treatment of ulcer with H₂ B can be prevented bycombining it with a gastrin receptor antagonist (see, Table 7).

Experimental results shown above demonstrate that the benzodiazepinederivatives of the present invention have antagonistic effect againstgastrin receptor and CCK-B receptor, and that a hybrid-type compoundconsisting of a compound of the present invention and an H₂ B isinhibitory against the acid-rebound phenomenon and decrease in theprotective function of gastric mucosa following anti-ulcer treatment,and is useful as an anti-ulcer agent unaccompanied by relapse.

The method of preparing pharmaceutical compositions of the presentinvention is shown below, which is provided only for illustrativepurpose.

    ______________________________________                                        Compound 17ix         50.0   mg                                               Lactose               128.0  mg                                               Potato starch         40.0   mg                                               Magnesium stearate    2.0    mg                                                                     220.0  mg                                               ______________________________________                                    

A 10% viscous solution of potato starch was prepared by heating.Compound 17ix, lactose and the rest of potato starch were mixed with theresultant viscous solution and granulated by passing through a sieve(mesh size 1.5 mm). The granules were dried at 45° C., passed throughthe same sieve again, mixed with magnesium stearate and formulated withtabletting machine.

    ______________________________________                                        Compound 17ix          200.0  mg                                              Lactose                120.0  mg                                              Corn starch            70.0   mg                                              Polyvinylpyrrolidone   8.0    mg                                              Magnesium stearate     2.0    mg                                                                     400.0  mg                                              ______________________________________                                    

Compound 17ix, lactose and corn starch were moistured homogeneously withaqueous polyvinylpyrrolidone solution and granulated by passing througha sieve (mesh size 2.0 mm). The granules were dried at 50° C. withcirculating air system, re-granulated by passing through a sieve (meshsize 1.5 mm), mixed with magnesium stearate, and formulated withtabletting machine.

Formulation 3 Capsules

Compound 17ix (60.0 mg) was pulverized to fine powder and filled intocapsules to give capsule formulation.

Formulation 4 Suppository

    ______________________________________                                        Compound 17ix        60.0    mg                                               Suppository base     1640.0  mg                                                                    1700.0  mg                                               ______________________________________                                    

Finely pulverized Compound 17ix was suspended into melted suppositorybase. The resultant suspension was cooled to 40° C. and poured into aslightly cooled suppository mold at 37° C.

Formulation 5 Suppository

    ______________________________________                                        Compound 17ix        200.0   mg                                               Suppository base     1500.0  mg                                                                    1700.0  mg                                               ______________________________________                                    

The suppository base was melted. An active ingredient pulverized at 38°C. was dispersed into the melted base homogeneously. The dispersion wascooled to 35° C. and poured into previously cooled suppository mold.

Formulation 6 Suspension

    ______________________________________                                        Compound 17ix          4.0    g                                               Carboxymethyl cellulose                                                                              0.1    g                                               Methyl p-hydroxybenzoate                                                                             0.05   g                                               Propyl p-hydroxybenzoate                                                                             0.01   g                                               Sucrose                10.0   g                                               Glycerin               5.0    g                                               70% Sorbitol solution  20.0   g                                               Fragrance              0.3    g                                               Distilled water                                                                                      1000   ml                                              ______________________________________                                    

Distilled water was warmed at 70° C.and methyl p-hydroxybenzoate, propylp-hydroxybenzoate, glycerin and carboxymethyl cellulose are dissolvedtherein. The resultant solution was cooled to room temperature. Compound17ix was added dropwise to the solution with stirring and dispersedhomogeneously. Sucrose, sorbitol and fragrance were added and dissolved.The resultant suspension was subjected to deaeration under vacuum withstirring.

What is claimed is:
 1. A compound of the formula (I): ##STR16## whereinR₁ is a bond, --CH₂ --, --CH₂ O--, --OCH₂ --, --SCH₂ -- or a group ofthe formula: ##STR17## R₂ is a lower alkyl, --COOR₅, --CONH(CH₂),COOR₅,--CONHSO₂ R₅, --SO₂ NHCOR₅, or a heterocyclic group selected from thegroup consisting of furyl, thienyl, tetrazolyl, pyrrolyl, pyrazolyl,imidazolyl, oxazolyl, thiazolyl pyridinyl, oxadinyl, triazinyl,pyrrolidinyl, thiazolidinyl, oxazolidinyl imidazolidinyl, thiazolinyl,oxazolinyl, imidazolinyl, piperidinyl, piperadinyl, morpholinyl,thiomorpholinyl, oxadiazolyl and dioxanyl, which is unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of hydroxy, carbonyl, amino, amino protected with anamino-protecting group, halogen, lower alkyl and lower alkoxy (R₅ is ahydrogen atom, lower alkyl or benzyl and n is an integer of 1 to 5); R₃is a bond, --CO-- or --CONH--; and R₄ isa heterocyclic group selectedfrom the group consisting of furyl, thienyl, tetrazolyl, pyrrolyl,pyrazolyl, imidazolyl, oxazolyl, thiazolyl, pyridinyl, oxadinyl,triazinyl, pyrrolidinyl, thiazolidinyl, oxazolidinyl, imidazolidinyl,thiazolinyl, oxazolinyl, imidazolinyl, piperidinyl, piperadinyl,morpholinyl, thiomorpholinyl, oxadiazolyl and dioxanyl, which isunsubstituted or substituted with one or more substituents selected fromthe group consisting of hydroxy, carbonyl, amino, amino protected withan amino-protecting group, halogen, lower alkyl and lower alkoxy, loweralkyl which is unsubstituted or substituted with one or moresubstituents selected from the group consisting of hydroxy, carbonyl,amino, amino protected with an amino-protecting group, halogen, loweralkyl and lower alkoxy, lower cycloalkyl, which is unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of hydroxy, carbonyl, amino, amino protected with anamino-protecting group, halogen, lower alkyl and lower alkoxy, arylselected from the group consisting of phenyl and naphthyl which isunsubstituted or substituted with one or more substituents selected fromthe group consisting of hydroxy, carbonyl, amino, amino protected withan amino-protecting group, halogen, lower alkyl and lower alkoxy, alower alkoxycarbonyl group, or a pharmaceutically acceptable saltthereof, and wherein R₃ is the same at each instance in said compound,and wherein R₄ is the same at each instance in said compound.
 2. Thecompound of the formula (I) of claim 1, wherein R₃ is --CO-- and R₄ is alower cycloalkyl group.
 3. The compound of claim 1 or 2, wherein R₁ -R₂is --COOR₅, --CONHSO₂ R₅, --SO₂ NHCOR₅, --CH₂ COOR₅, --OCH₂ COOR₅, -SCH₂ COOR₅, tetrazolylmethyloxy or a 5-membered heterocyclic groupcontaining a N atom.
 4. A compound of the formula (III): ##STR18##wherein R₁ is a bond, --CH₂ --, --CH₂ O--, --OCH₂ --, --SCH₂ -- or agroup of the formula: ##STR19## and R₂ is a lower alkyl, --COOR₅,--CONH(CH₂),COOR₅, --CONHSO₂ R₅, --SO₂ NHCOR₅, or a heterocyclic groupselected from the group consisting of furyl, thienyl, tetrazolyl,pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, pyridinyl,oxadinyl, triazinyl, pyrrolidinyl, thiazolidinyl, oxazolidinyl,imidazolidinyl thiazolinyl, oxazolinyl, imidazolinyl, piperidinyl,piperadinyl, morpholinyl, thiomorpholinyl, oxadiazolyl and dioxanyl,which is unsubstituted or substituted with one or more substituentsselected from the group consisting of hydroxy, carbonyl, amino, aminoprotected with an amino-protecting group, halogen, lower alkyl and loweralkoxy (R₅ is a hydrogen atom, lower alkyl or benzyl and n is an integerof 1 to 5).
 5. A pharmaceutical composition comprising a therapeuticallyeffective amount of at least one compound of claim 1 and at least onepharmaceutically acceptable carrier therefor.